2009年 · 第122卷第09期

Abstract：Background Neurocysticercosis is the infection of the nervous system by the larvae of Taenia solium (T. solium). Despite continuous effort, the experimental diagnosis of neurocysticercosis remains unresolved. Since the cerebrospinal fluid (CSF) contacts with the brain, dynamic information about pathological processes of the brain is likely to be reflected in CSF. Therefore, CSF may serve as a rich source of putative biomarkers related to neurocysticercosis. Comparative proteomic analysis of CSF of neurocysticercosis patients and control subjects may find differentially expressed proteins. Methods Two-dimensional difference in gel electrophoresis (2D-DIGE) was used to investigate differentially expressed proteins in CSF of patients with neurocysticercosis by comparing the protein profile of CSF from neurocysticercosis patients with that from control subjects. The differentially expressed spots/proteins were recognized with matrix-assisted laser desorption/ionization-time of flight-time of flight (MALDI-TOF-TOF) mass spectrometry. Results Forty-four enzyme digested peptides were obtained from 4 neurocysticercotic patients. Twenty-three were identified through search of the NCBI protein database with Mascot software, showing 19 up-expressed and 4 down-expressed. Of these proteins, 26S proteosome related to ATP- and ubiquitin-dependent degradation of proteins and lipocalin type prostaglandin D synthase involved in PGD2-synthesis and extracellular transporter activities were up-expressed, while transferrin related to iron metabolism within the brain was down-expressed. Conclusions This study established the proteomic profile of pooled CSF from 4 patients with neurocysticercosis, suggesting the potential value of proteomic analysis for the study of candidate biomarkers involved in the diagnosis or pathogenesis of neurocysticercosis。

Abstract：Background Significant progress has taken place over the past 50 years in the control of schistosomiasis japonica in China. However, the available data suggested that schistosomiasis has re-emerged shortly after the World Bank Loan Project which was conducted from 1992 to 2001. The national control program with a revised strategy to control schistosomiasis by using integrated measures has been implemented since 2005. In this study, we aimed to evaluate the effect of the national program on schistosomiasis control from 2005 to 2008.Methods A retrospective study was carried out to analyze the epidemic patterns of acute infections with Schistosoma japonicum (S. japonicum), based on the number of acute cases annually collected from the web-based national communicable diseases reporting system from 2005 to 2008.Results A total of 564, 207, 83 and 57 acute cases infected with S. japonicum were reported nationwide in 2005, 2006, 2007 and 2008, respectively, with an average annual reduction rate of 46.35% during last four years. Six outbreaks of acute infection with S. japonicum were reported in 2005 but none in the period of 2006 to 2008. All acute cases that were reported mainly came from the lake regions and became infected during the higher risk periods from the 27th to 43rd weeks of the year. Most of these cases are students (44.87%), farmers (31.51%) and fishermen (7.79%) who got the infection by water contact mainly through swimming (41.49%) and production activities (40.25%). With time, the proportion of imported cases among all acute cases increased due to more frequent movement of people that has occurred with a more mobile population.Conclusions The national control program on schistosomiasis aliened with the revised control strategy has been effectively brought into effect. However, there is still a significant risk of infection among students, farmers and fishermen living in the lake regions. Therefore, it is important to strengthen control measures among risk populations in the high risk areas of transmission, or the lake regions。

Abstract：Background There are no reports on the association between perinatal characteristics and comprehensive physical function in Chinese elderly people. In this study, we traced 875 subjects who were born at the Peking Union Medical College Hospital (PUMCH) of China from 1921 to 1941. The purpose of this study was to determine the effects of perinatal characteristics on activities of daily living (ADL) function in the geriatric period. Methods Birth data of 875 subjects were obtained from obstetric birth records of PUMCH. Adulthood data collection was conducted in the outpatient clinics of PUMCH. During the clinic visits, trained research staff administered physical examinations, activities of daily living scale and a demographic questionnaire. ADLs of all subjects were assessed with the activities of daily living scale.Results There were 101 subjects whose ADL function was limited and the rate of ADL limitation was 11.5%. Binary logistic regression analyses results showed that the main influencing factors of ADL were age, maternal age at birth, occupation, daily exercise and chronic disease. Subjects whose maternal age at their birth exceeded 35 years were at 2.202 times (1.188-4.083) greater risk of ADL limitation when we applied multivariate logistic regression models. Conclusions This study validated the relationship between perinatal characteristics and ADL in the geriatric period. An older maternal age at birth could predict a higher ADL limitation rate in the geriatric period。

Abstract：Background The optimal timing to start continuous renal replacement therapy (CRRT) for acute kidney injury (AKI) patients has not been accurately established. The recently proposed risk, injury, failure, loss, end-stage kidney disease (RIFLE) criteria for diagnosis and classification of AKI may provide a method for clinicians to decide the "optimal timing" for starting CRRT under uniform guidelines. The present study aimed: (1) to analyze the correlation between RIFLE stage at the start of CRRT and 90-day survival rate after CRRT start, (2) to further investigate the correlation of RIFLE stage with the malignant kidney outcome in the 90-day survivors, and (3) to determine the influence of the timing of CRRT defined by RIFLE classification on the 90-day survival and malignant kidney outcome in 90-day survivors. Methods A retrospective cohort analysis was performed on the data of 106 critically ill patients with AKI, treated with CRRT during a 6-year period in a university affiliated surgical intensive care unit (SICU). Information such as sex, age, RIFLE stage, sepsis, sepsis-related organ failure assessment (SOFA) score, number of organ failures before CRRT, CRRT time during SiCU, survival, and kidney outcome conditions at 90 days after CRRT start was collected. According to their baseline severity of AKI at the start of CRRT, the patients were assigned to three groups according to the increasing severity of RIFLE stages: RIFLE-R (risk of renal dysfunction, R), RIFLE-I (injury to the kidney, I) and RIFLE-F (failure of kidney function, F) using RIFLE criteria. The malignant kidney outcome was classified as RIFLE-L (loss of kidney function, L) or RIFLE-E (end-stage kidney disease, E) using RIFLE criteria. The correlation between RIFLE stage and 90-day survival rate was analyzed among these three RIFLE-categorized groups. Additionally, the association between RIFLE stage and the malignant kidney outcome (RIFLE-L+RIFLF-E) in the 90-day survivors was analyzed.Results Fifty-three of the overall 106 patients survived to 90 days after the start of CRRT. There were 16, 22 and 68 patients in RIFLE-R, RIFLE-I and RIFLE-F groups respectively with corresponding 90-day survival rate of 75.0% (12/16), 63.6% (14/22) and 39.7% (27/68) (P<0.01, compared among groups). The percentage of the malignant kidney outcome of 90-day survivors in the RIFLE-R, RIFLE-I, and RIFLE-F groups was 16.7% (2/12), 21.4% (3/14) and 55.6% (15/27), respectively (P for trend <0.01). After adjustment for other baseline risk factors, the relative risk (RR) for the 90-day mortality significantly increased with baseline RIFLE stage. Patients in RIFLE-F had a higher RR of 1.96 (95% confidence interval (CI): 1.06-3.62) than patients in RIFLE-I (RR: 1.09, 95% CI: 0.55-2.15) compared with patients in RIFLE-R (P for trend <0.01). Similarly, baseline RIFLE stage also significantly correlated with the odds ratio (OR) for the malignant kidney outcome in 90-day survivors (P for trend <0.05). Ninety-day survivors in the RIFLE-F group had a borderline significantly highest OR of 6.88 (95% CI: 0.85-55.67).Conclusions The RIFLE classification may be used to predict 90-day survival after starting CRRT and the malignant kidney outcome of 90-day survivors in the critically ill patients with AKI treated with CRRT. Starting CRRT prior to RIFLE-F stage may be the optimal timing. Prospective, multi-center, randomized controlled trials are needed to confirm its predictive value in these patients。

Abstract：Background Non-small cell lung cancer (NSCLC) is the leading cause of cancer mortality worldwide. Platelet activation may play an important role in pathologic progress in lung cancer. In this study, we aimed to clarify the influence of activated platelets on lung cancer generation and growth, and the relationship among these functional and ultrastructural changes of platelets and the severity of pathogenetic condition in these patients with NSCLC.Methods One hundred and thirty-six cases of patients with pathologically confirmed NSCLC were included in this study. Fifty-four healthy people were enrolled as controls. The change of ultra microstructure and activity of blood platelets were observed under the transmission and scanning electron microscope. Simultaneous determination of plasma granule membrane protein 140 (GMP-140) was made.Results Transmission electron microscopy showed remarkable changes of ultra microstructure of platelets in patients with NSCLC, including swelling, increase of o-granules, vesicles, and glycogenosome. Scanning electron microscopy showed many more surface processes and wrinkles on platelets in patients with NSCLC. The reference plasma levels of GMP-140 of healthy controls were (18.2±2.7) μg/L. The plasma levels of GMP-140 in patients with NSCLC were (47.8±12.3) μg/L, which were much higher than those of the controls. There was a medium positive correlation between plasma levels of GMP-140 and amount of o-granules (r=0.514, P<0.01) and a high positive correlation between plasma levels of GMP-140 and area of platelet (r=0.84, P<0.01) in patients with NSCLC. The Kaplan-Meier survival curve analysis showed significant shift to the left in patients with NSCLC whose α-granules per platelet were 19 or more compared to those 18 or less (Log rank statistic, X2=17.38, P <0.01).Conclusions There are significant activated changes of ultra microstructure and increased activity of blood platelets in patients with NSCLC. These activated platelets may play an important role in the generation and growth of lung cancer. These changes can be used as a diagnostic index of severity, progression, and prognosis of NSCLC。

Abstract：Backgroud Little is known about the influence of liver transplantation on the pharmacokinetics of most anesthetic drugs. The goal of this study was to study the population pharmacokinetics of remifentanil in the different phases of orthotopic liver transplantation (OLT) and the influence of relevant factors.Methods Thirteen adult patients undergoing OLT were enrolled. A single bolus infusion of remifentanil 5 μg/kg was administered during the preanhepatic, anhepatic and neohepatic phases of OLT. Arterial blood samples of 1.5 ml were collected at 0 (baseline), 1, 2, 3, 5, 7, 10, 15, 20, 25, 30, 45, 60 and 90 minutes after drug administration. Remifentanil concentration was assayed by high-performance liquid chromatography/mass spectrometry/mass spectrometry (HPLC/MS/MS). Population pharmacokinetic modeling was performed using nonlinear mixed-effects modeling (NONMEM).Results The pharmacokinetics of remifentanil in patients undergoing OLT was best described by a two-compartment open model. The pharmacokinetic parameters were not influenced by age, gender, operative phase, blood temperature, rehydration volume, or blood loss volume during sampling. The volume of distribution in the central compartment (V1) and the volume of distribution in the peripheral compartment (V2) were influenced by body weight. Conclusions The population pharmacokinetics of remifentanil in patients undergoing OLT can be well described by a two-compartment open model. The functional status of the liver does not significantly affect the pharmacokinetics of remifentanil, but the body weight is an influential factor of V1 and V2。

Abstract：Background Arthritogenic T lymphocytes with common T cell receptor (TCR) Vβ clonotypes, infiltrating in the articulars of rheumatoid arthritis (RA) patients, play a central role in the pathogenesis of RA. TCR Vβ5.2 and TCR Vβ8.2 are the main pathogenic T cell clonotypes in the course of collagen-induced arthritis (CIA) progression in Lewis rats. To investigate a TCR-based immunotherapy for RA, we constructed recombinant DNA vaccines encoding TCR Vβ5.2 and TCR Vβ8.2, and evaluated the inhibitive effects of the two vaccines on CIA rats. Methods Genes encoding TCR Vβ5.2 and TCR Vβ8.2 were amplified by RT-PCR from spleen lymphocytes of Lewis rats and cloned into the eukaryotic expression vector pTargeT. The expression of vaccines was confirmed by RT-PCR and immunohistochemistry. The inhibitive effects of the vaccines on articulars of CIA rats were assessed with arthritis index evaluation and histology. Interferon γ (IFN-γ) and interleukin (IL)-4 production by spleen lymphocytes were tested with enzyme-linked immunospot assay (ELISPOT) technique, the changes in peripheral CD4+ and CD8+ lymphocyte populations were tested by flow cytometry, and the level of anti-CII antibody in serum was assayed by enzyme-linked immunosorbent assay (ELISA).Results Recombinant DNA vaccines pTargeToTCR Vβ5.2 and pTargeT-pTCR Vβ8.2 were successfully constructed. Both vaccines inhibited CIA, which alleviated the arthritis index score (P<0.05), decreased the level of IFN-γ (P<0.05), and reduced the ratio of CD4+/CD8+ lymphocytes (P<0.05) and the anti-CII antibody in serum (P<0.05). In addition, the histological change in DNA-vaccinated rats was less serious than CIA rats. Compared to pTCR Vβ 8.2 and pTCR Vβ5.2 groups, the group that was injected with a combination of the two vaccines showed stronger inhibitive effects on CIA than either individual vaccine.Conclusion The recombinant plasmids pTargeT-TCR Vβ5.2 and pTargeT-TCR Vβ8.2 have obvious inhibatory effects on CIA rats and better effects could be achieved when the vaccines were used in combination。

Abstract：Background Ca2+in the central nervous system plays important roles in brain physiology, including neuronal survival and regeneration in rats with injured facial motoneurons. The present research was to study the modulations of intracellular free Ca2+ concentrations by cholinergic receptors in rat facial nucleus, and the mechanisms of the modulations. Methods The fluorescence intensity of facial nucleus in Fluo-3 AM loaded acute brainstem slices was detected by applying intracellular free Ca2+ measurement technique via confocal laser scanning microscope. The changes of fluorescence intensity of facial nucleus indicate the average changes of intracellular free Ca2+ levels of the neurons. Results Acetylcholine was effective at increasing the fluorescence intensity of facial nucleus. Muscarine chlorlde induced a marked increase of fluorescence intensity in a concentration dependent fashion. The enhancement of fluorescence intensity by muscarine chloride was significantly reduced by thapsigargin (depletor of intracellular Ca2+ store; P<0.01), rather than Ca2+ free artifical cerebrospinal fluid or EGTA (free Ca2+ chelator; P>0.05). And the increase of fluorescence intensity was also significantly inhibited by pirenzepine (M1 subtype selective antagonist; P<0.01) and 4-DAMP (M3 subtype selective antagonist; P<0.01). In addition, fluorescence intensity was markedly increased by nicotine. The enhancement of fluorescence intensity by nicotine was significantly reduced by EGTA, nifedipine (L-type voltage-gated Ca2+ channel blocker), dihydro-β-erythroidine (α4β2 subtype selective antagonist), and in Ca2+ free artificial cerebrospinal fluid (P<0.01), but not in the presence of mibefradil (M-type voltage-gated Ca2+ channel blocker) or thapsigargin (P>0.05).Conclusions The data provide the evidence that muscarinic receptors may induce the increase of intracellular free Ca2+ levels through the Ca2+ release of intracellular Ca2+ stores, in a manner related to M1 and M3 subtypes of muscarinic receptors in rat facial nucleus. Nicotine may increase intracellular free Ca2+ concentrations via the influx of extracellular Ca2+ mainly across L-type voltage-gated Ca2+ channels, in a manner related to the α4β2 subtype of nicotinic receptors。

Abstract：Background Myoglobin is expressed exclusively in striated skeletal muscles and has been implicated in nitric oxide scavenging. Accumulating data suggest a critical role for nitric oxide in both the endogenous and therapeutic angiogenic response to ischemia. A clear role for myoglobin in ischemic skeletal muscle is uncertain. We hypothesized that myoglobin overexpression has an adverse impact on the angiogenic response to ischemia.Methods Muscle-specific myoglobin over-expressing transgenic mice (MbTG, n=11), wild type littermates (WT, n=23) underwent unilateral femoral artery ligation and excision. Laser doppler perfusion imaging was used to monitor changes in hindlimb perfusion before surgery and weekly after surgery up to 28 days. Tissue ischemia was assessed by a necrosis incidence. Upon termination of the experiment (28 days after surgery), skeletal muscles (gastrocnemius, and tibialis anterior) were harvested, the distal part of the muscle was frozen and embedded for histology study, the proximal part was used either to detect vascular endothelial growth factor (VEGF) level with enzyme-linked immunosorbent assays (ELISA) or to determine the proliferation (proliferating call nuclear antigen (PCNA)) and apoptosis (Bax, and Bcl-2) condition in ischemic muscle by Western blotting. Capillaries were stained with endothelial phosphate alkaline staining and vascular density was expressed in capillaries/fiber.Results The recovery of perfusion in MbTG mice was similar to that of WT mice on day 7 (0.485±0.095 vs 0.500±0.084)but was significantly less on day 14 (0.536±0.086 vs 0.623±0.077, P <0.05), day 21 (0.588±0.082 vs 0.684±0.068, P <0.01) and day 28 (0.606±0.079 VS 0.733±0.093, P<0.01). The necrosis incidence was higher in MbTG than in WT (54.5% vs 21.6%). Vascular density was less in MbTG compared with that in WT (gastrocnemius 0.19±0.08 vs 0.30±0.08, P <0.05; tibialis anterior 0.22±0.11 vs 0.33±0.04, P<0.05). With ischemic injury, the VEGF level was increased in both MbTG and WT (45.2% and 20.4%, respectively). Western blotting showed that after hindlimb ischemia the proliferation was similar in both MbTG and WT, however, apoptosis was increased in MbTG relative to WT, shown as more expression of Bax and less expression of Bcl-2.Conclusion An increase in expression of myoglobin protein in skeletal muscle reduces the endogenous perfusion recovery following surgically induced hind-limb ischemia。

Abstract：Background Tyrosine kinase signaling cascades play a critical role in the pathogenesis of allergic airway inflammation. Sunitinib, a multitargeted receptor tyrosine kinase inhibitor, has been reported to exert potent immunoregulatory, anti-inflammatory and anti-fibrosis effects. We investigated whether sunitinib could suppress the progression of airway inflammation, airway hyperresponsiveness (AHR), and airway remodeling in a murine model of chronic asthma. Methods Ovalbumin (OVA)-sensitized mice were chronically challenged with aerosolized OVA for 8 weeks. Some mice were intragastrically administered with sunitinib (40 mg/kg) daily during the period of OVA challenge. Twelve hours after the last OVA challenge, mice were evaluated for the development of airway inflammation, AHR and airway remodeling. The levels of total serum immunoglobulin E (IgE) and Th2 cytokines (interieukin (IL)-4 and IL-13) in bronchoalveolar lavage fluid (BALF) were measured by ELISA. The expression of phosphorylated c-kit protein in the lungs was detected by immunoprecipitation/Western blotting (IP/WB) analysis.Results Sunitinib significantly inhibited eosinophilic airway inflammation, persistent AHR and airway remodeling in chronic experimental asthma. It reduced levels of total serum IgE and BALF Th2 cytokines and also lowered the expression of phosphorylated c-kit protein in remodelled airways.Conclusions Sunitinib may inhibit the development of airway inflammation, AHR and airway remodeling. It is potentially beneficial to the prevention or treatment of asthma。

Abstract：Background Galectin-3 is the most recently identified advanced glycosylation end products (AGEs) binding protein. This study aimed to investigate the effects of AGEs and rosiglitazone on the expression and secretion of galectin-3 in cultured human renal mesangial cells (HRMCs).Methods HRMCs were incubated with different concentrations of AGE-bovine serum albumin (BSA) (0, 50, 100, 200, and 400 mg/L) for different time (0, 24, 36, 48, and 72 hours), and exposed to AGE-BSA in the presence of different concentrations of rosiglitazone (1, 10, and 100 μmol/L). The mRNA and protein expression of galectin-3 in HRMCs were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Westem blotting. The culture medium of HRMCs was collected and concentrated, and the content of galectin-3 in the medium was detected by Western blotting. Results Both RT-PCR and Western blotting revealed that AGE-BSA up-regulated the expression of galectin-3 in HRMCs in a concentration- (P<0.05) and time-dependent (P<0.05) manner compared with the control. Compared with the control, AGE-BSA elevated the content of galectin-3 in the culture medium of HRMCs time- and concentration-dependently (P<0.05, respectively). Both protein and mRNA expression of galectin-3, and its content in the medium of HRMCs exposed to different concentrations of rosiglitazone in the presence of AGE-BSA were increased compared with those of cells exposed to AGE-BSA alone (P<0.05). Rosiglitazone increased the expression and secretion of galectin-3 in a dose-dependent manner (P<0.05).Conclusions AGEs up-regulates the expression and secretion of galectin-3 in HRMCs. Rosiglitazone further enhances the upregulation of galectin-3 in HRMCs induced by AGEs, which suggests that rosiglitazone may play a role of reno-protection via up-regulation of galectin-3。

Abstract：Background Equatorial lens epithelial cells proliferate and differentiate into fiber cells throughout life, while central lens epithelial cells proliferate little and do not form fiber cells. This study aimed to investigate the differences in gene expression between the central and the peripheral epithelial cells of the bovine lens. Methods Lens epithelia were dissected into central (≤11.5 mm diameter, cLEC) and peripheral regions (pLEC). The differences in gene expression and protein accumulation between these two regions were assayed by microarray analysis and two-dimensional electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Differently expressed proteins were validated by immunoanalyses. Results By microarray analysis, 67 transcripts were at least two-fold lower and 269 at least two-fold higher in pLEC compared with that in cLEC. Thirty-four protein spots, including 20 in cLEC and 14 in pLEC, were identified by two dimensional electrophoresis and mass spectrometry. Of these 34 protein products, 28 were represented by probe sets on the microarray. Nine transcripts changed in the same direction and four transcripts in the opposite direction to their protein products. Immunoanalyses revealed that three (mitogen-activated protein kinase 1 (MAPK1), nidogen (NID), small nuclear ribonucleoprotein N (SNRPN)) out of four transcripts with opposite change between 2-DE and microarray assay showed the same changes as the results of 2-DE gel analyses. The genes differently expressed between cLEC and pLEC mainly include those related to the MAPK, transforming growth factor β (TGFβ) signaling and glycolysis pathways.Conclusion The results suggested that there were distinctly different genome activities, including a specific group of pathways, between central and peripheral lens epithelial cells。

Abstract：Background Virus with nucleoside reverse transcriptase inhibitors (NRTIs) or nonnucleoside reverse transcdptase inhibitors (NNRTIs) resistant mutations show different evolution tendencies when the anti-viral therapies are interrupted. Understanding the replication fitness of drug-resistant virus is important for the study of the prevalence of drug-resistance. For this purpose, we characterized the replication capacity of HIV-1 virus carrying lamivudine (3TC) or nevirapine (NVP) resistant mutations.Methods 3TC and NVP resistant variants were induced in vitro by selecting wild type virus in the presence of drugs. For the competitive replication assay, drug-resistant variants were cocultured with wild-type virus in the presence or absence of drugs. The ratios of the viral species were determined over time by using a real-time RT-PCR-based assay. Results 3TC-resistant (M184I mutation) and NVP-resistant (Y181I mutation) virus should be selected in vitro in two different ways. The competitive replication assay showed that the ratio of virus carrying a M184I mutation increased from 98.8%, while the wild type virus decreased to 1.2% after 4 passages in the presence of 3TC; the percentage of virus carrying the Y181I mutation increased to 90.5%, while wild type virus decreased to 9.5% in the presence of NVP. In the absence of drugs, the ratio of virus carrying the M184I mutation decreased to 5.3%, while wild type virus increased to 94.7%; the ratio of virus carrying Y181I increased to 75%, while wild type virus decreased to 25% after 4 passages.Conclusions The NVP-resistant virus is fitter than wild type virus even in the absence of NVP that may be the reason that NNRTIs-resistant virus is spreading quickly。

Abstract：Background Continent cutaneous diversion (CCD) has been widely used in almost any lower urinary reconstruction. We have been continually trying to modify this procedure because of the high complications rate, especially as they relate to the efferent tube. In this study, we reported a modified procedure with a tapered ileum wrapped by the rectus abdominalis flap (RAMF) and assessed the feasibility of this new technique to achieve urinary continence. Methods A procedure in which two ileal segments were tapered and connected to a U-shaped reservoir was performed in ten dogs. A RAMF with its blood supply was wrapped around one of the tapered ileum, in control groups, the tapered ileum was brought to the abdominal skin. Urodynamic studies were conducted In the 1st, 3rd and 6th months post-operatively. The data of maximum inner pressure (MIP) and functional pressure length (FPL) in every group at each phase were recorded. Retrograde radiograms of the efferent limbs were performed before sacrifice.Results MIP in the study group was significantly higher than that in the control group at each phase (P<0.05). However, no significant differences in MIP or FPL were found in the study group between an empty and full reservoir. In the control group, MIP increased (P<0.05) and FPL decreased significantly (P<0.05) compared with an empty and full reservoir. Retrograde radiograms confirmed that efferent limbs were positioned straigh beneath the abdominal wall. Histological examination of the study group demonstrated a layer of striated muscle around the outside surface of the ileum.Conclusion The continent mechanism of tapered ileum can be enhanced by extra support from wrapped RAMF。

Abstract：Backgroud AmpC β-lactamases and extended-spectrum β-lactamases (ESBLs) are becoming predominant causes of resistance to third and forth-generation cephalosporins in Klebsiella pneumoniae (K. Pneumoniae). It is very difficult to treat infectious diseases caused by multidrug-resistant K. Pneumoniae. The purpose of the present study was to investigate transconjugation and characteristics of β-lactamase genes in K. Pneumoniae producing AmpC β-lactamases and ESBLs.Methods AmpC β-lactamases were detected by three-dimension test and ESBLs by disc confirmatory test. Minimum inhibitory concentrations (MICs) were determined by agar dilution. Transfer of resistance to EC600 (Rif') was attempted by conjugation in broth and screened on agar containing cefotaxime (2 μg/ml) plus rifampin (1024 μg/ml). The genes encoding AmpC or ESBLs and their transconjugants were detected by PCR and verified by DNA sequencing. Results The resistant rates to ampicillin and piperacillin were 100% in 18 isolates of K. Pneumoniae. However, imipenem was still of great bactericidal activity on K. Pneumoniae, and its MIC50 was 0.5 μg/mL. Eleven β-lactamase genes, including TEM-1, TEM-11, SHV-13, SHV-28, CTX-M-9, CTX-M-22, CTX-M-55, OXA-1, LEN, OKP-6 and DHA-1, were found from 18 isolates. And at least one β-lactamase gene occurred in each isolate. To our surprise, there were six β-lactamase genes in the CZ04 strain. Among 18 isolates of K. Pneumoniae, the partial resistant genes in 8 isolates were conjugated successfully, which had 100% homological sequence with donors by sequence analysis. Compared with donors, 8 transconjugants had attained resistance to most β-lactams, including ampicillin, piperacillin, cefoxitin, cefotaxime and aztreonam, or even amikacin and gentamicin.Conclusions R plasmids can be easily transferred between the resistant and sensitive negative bacilli. It is very difficult to block and prevent the spread of antimicrobial resistance. So more attention should be paid to reducing the frequency, times and dosage of antimicrobials, especially third or fourth cephalosporins。

Abstract：Background The autosomal dominant form of retinitis pigmentosa (ADRP) can be caused by mutations in 14 genes and further loci remains to be identified. This study was intended to identify mutations in a Chinese pedigree with ADRP. Methods A large Chinese family with retinitis pigmentosa was collected. The genetic analysis of the family suggested an autosomal dominant pattern. Microsatellite (STR) markers tightly linked to genes known to be responsible for ADRP were selected for linkage analysis. Exons along with adjacent splice junctions of PRPF31 were amplified by polymerase chain reaction (PCR) and screened by direct sequencing.Results The caused gene of ADRP was mapped to 19q13.4 between markers D19S572 and D19S877, with a maximum LOD score of 3.01 at marker D19S418 (recombination fraction=0).Conclusion The affected gene linked to the 19q13.4 in a Chinese family with ADRP, which is different from other mutations at the same loci in other Chinese families。

During clinical work, some side-effects may occur to patients, part of which are caused by the specific pharmacological effects of drugs and some of which are non-specific. Although these phenomena happen from time to time, burdening the anguish and expenditure of patients, their nature is still less understood. Recently, as the research of the placebo effect become deeper and deeper, clinicians and researchers have gradually realized that mind plays an important role in the occurrence of non-specific side-effects, which is called "nocebo effect" professionally, the evil side of placebo effect. This article would expatiate on nocebo effect in detail from several aspects, such as its mechanism, effect, influencing factors and discuss how to make it known and treated in clinical practice and clinical trials。

The long arm of human chromosome 12 contains a region that has been found to be amplified in a number of different tumors, including osteosarcomas and soft tissue sarcomas,1 There are more than 5 genes located in this area such as CDK2, CDK4, WNT1, MDM2 and WNTIOb.1 CDK4 gene consists of eight exons, of which the start codon is located in the beginning of exon 2 and the stop codon in the beginning of exon 8. CDK4 is a member of the Ser-Thr protein kinase family and its catalytic domain extends from amino acid 6 to 295. CDK4 is expressed in a variety of normal cells and tissues as well as in cancer cells. CDK4 constitutes the catalytic subunit of a heterodimeric Ser/Thr protein kinase that is involved in controlling progression through the G1 phase of the cell cycle. Germ-line mutations in the CDK4 gene have so far only been found in families with inherited malignant melanoma and multiple atypical nevi. The mutations affect the Arg encoded by codon 24, changing it either to Cys or to His。

Chronic obstructive pulmonary disease (COPD) is a major cause of chronic morbidity and mortality throughout the world. It is a preventable and treatable disease with significant extra pulmonary effects that may contribute to the severity in individual patients. It is characterized by airflow limitation that is not fully reversible,1 The diagnosis of COPD should be confirmed by spirometry. Body mass index (BMI) is an important indicator that can well reflect nutritional status of patients, and low BMI is an independent risk factor for mortality in patients with COPD.2

Antithyroid drugs are molecules known as thionamides that inhibit thyroid hormone synthesis by interfering with thyroid peroxidase mediated iodination of tyrosine residues in thyroglobulin. These extensively used drugs are associated with a variety of well-known side effects such as anti-neutrophil cytoplasmic antibody (ANCA)-positive vasctilitis, granulocytopenia and aplastic anemia。

The cases of aberrant retroesophageal right subclavian artery (ARRSA) with esophageal cancer are rare. The treatment could be a challenge. We present a case of ARRSA with esophageal cancer infiltration and a successful surgical treatment。

Although catheterization of the subclavian vein is a common procedure, it might be associated with life-threatening complications including accidental cannulation of subclavian artery.1-3 Rash sheath removal could result in fatal hemorrhage. We report a case of inadvertent 6F sheath cannulation of the right subclavian artery during a radiofrequency catheter ablation procedure and successful repair of the cannulation by Angio-Seal STS plus (St. Jude Medical, St. Paul, MN, US), a collagen-base vascular closure device。

To the Editor: We read the article by Ke et al1 with great interest, in which they investigated the usage of aspirin for the secondary prevention of ischemic stroke. The incidence of ischemic stroke and transient ischemic attack assessed by onset of clinical symptoms exhibits a marked circadian variation with a peak period during the morning. Stroke usually occurs unexpectedly or more frequently in the morning hours, between 7-12 a.m. In this morning period there is a higher aggregability of thrombocytes. Patients usually take aspirin in the morning for prevention as the treatment regimen is one tablet per day to be swallowed without chewing at least 30 minutes before breakfast (Figure). The highest plasma level of the drug occurs after the morning peak-incidence of the thromboembolic event, suggesting lower prophylactic effect of aspirin. Taking aspirin in the morning has its highest protective effect during the day, when normal physical activity exerts a protective action. Furthermore, this method of daily aspirin administration has its lowest protective value against cardio- and cerebrovascular events during the night and early rooming, when the lack of physical activity further augment the cascade of haemorheological events favoring platelet aggregation and subsequent ischemia.2-5

To the Editor: I read with great interest the recent article from the Sino-Global Registry of Acute Coronary Events (GRACE) of patients in China.1 This study includes patients from different parts of China, e.g., west (Xinjiang Autonomous Region Hospital), north (Harbin Medical University Affiliated First Hospital), east (Shanghai Medical University Affiliated 9th Hospital), and south (Guangdong Provincial Cardiovascular Institute). Therefore it is truly a very representative sample for a country as large and diversified as China。