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Chia (Salvia hispanica), originated in Mexico, has outstanding nutritional and health-promoting values, but it is the only ancient American Indian staple crop failed in introduction to the Old Continents. After the rediscovery and revival of chia as a new crop in recent years, the short-day (SD) habit is a crucial limitation for its worldwide cultivation. The circadian oscillator GIGNATEA (GI) is an important transcription factor regulating flowering time through photoperiod-pathway. In this study, we cloned the full-length cDNAs of two GI genes from chia, and analyzed the molecular characteristics of the genes and the encoded proteins. Alternative transcription initiation sites, alternative poly A tailing sites, and 5’-UTR intron retention exist in some of their mRNAs. The origin of GI gene accompanied with the transition from aquatic to terrestrial habits during plant evolution. GI duplication events occurred at order, family and genus levels in angiosperms. ShGI-1 and ShGI-2 were similar to each other in organ specificity with peak expression in small buds. In mature leaf, ShGI-2 is dominant over ShGI-1 in terms of expression level with highest expression in the afternoon, but on the Autumnal Equinox day ShGI-1 is dominant over ShGI-2 with peaks at noon and in the evening. KT, BR, GA3 and IAA upregulated the expression of ShGI-1 and ShGI-2 in long-days (LDs) and inhibited their expression in SDs, with GA3 being the most effective phytohormone. Under most abiotic stresses, ShGI expression fluctuated and returned to near-basal levels. ShGI expression was upregulated by low temperature. SA sharply upregulated ShGI expression after 24 h of treatment. This is the first report of GI genes from the order Lamiales, which will promote the dissection of flowering mechanism of chia and other Lamiales plants, enrich the evolution and expression characteristics of plant GIs, and promote the study on interaction between photoperiod and hormone pathways in flowering time control. © 2021 Friends Science Publishers

Sex differences in the long-term prognosis of symptomatic hypertrophic obstructive cardiomyopathy (HOCM) patients undergoing alcohol septal ablation (ASA) remain unclear, especially in the Chinese Han population.This cohort study included 320 HOCM Chinese Han patients who underwent ASA because of symptomatic left ventricular outflow tract (LVOT) obstruction. Patients were grouped according to sex: females (mean±standard deviation age [SD] 50.7±6.8 years) and males (mean±SD age 52.6±7.3 years). Individuals were followed over the long term.Pre-procedure, women had more symptoms (New York Heart Association [NYHA] class III-IV 67.3% vs 56.3%, p=0.03), more atrial fibrillation (23.5% vs 14.6%, p=0.047) than men. Transient complete atrioventricular block after ASA was more common in woman than in men (34.0 vs 23.4%; p=0.048). Residual LVOT gradient, post-procedural residual left ventricular wall thickness, NYHA functional class, and adverse arrhythmic events were comparable between the two groups. The 10-year survival rate (77% vs 89%, p=0.037) and the annual adverse arrhythmic event rate (1.3% vs 0.4%, p0.01) following ASA were significantly worse in women compared with men. Kaplan-Meier analysis showed a significantly lower survival in women compared with men (p=0.023). In multivariable modelling, female sex remained independently associated with higher all-cause mortality (hazard ratio, 1.12; 95% confidence interval, 1.08-1.27; p=0.03) when adjusted for age, NYHA class III-IV symptoms, and other cardiovascular comorbidities.Female patients with HOCM undergoing ASA tended to have more severe symptoms and adverse arrhythmic events. The 10-year survival rate after ASA was significantly worse in women compared with men with HOCM. Sex may need to be considered as an important factor in the clinical management of patients with symptomatic HOCM.

Yong Wang, Xiao-jiao Zhang, Hong-wei Zhao, Chengfu Wang, Defeng Luo, Qingkun Meng, Yu Zhu, Jie Tao, Baojun Chen, Yi Li, Aijie Hou, Bo Luan Department of Cardiology, The People’s Hospital of China Medical University, The People’s Hospital of Liaoning Province, Shenyang City, Liaoning Province 110016, People’s Republic of ChinaCorrespondence: Bo Luan; Aijie HouDepartment of Cardiology, The People’s Hospital of China Medical University, The People’s Hospital of Liaoning Province, No. 33, Wenyi Road, Shenhe District, Shenyang City, Liaoning Province 110016, People’s Republic of ChinaTel +86-17702488970Email luanbo2016@163.com; 1758624242@qq.comBackground: Retrograde microcatheter collateral channel (CC) tracking after successful wiring of septal CC is crucial for retrograde revascularization of coronary chronic total occlusion (CTO). However, the incidence, predictors, and strategies for failure of retrograde microcatheter CC tracking after successful wiring of septal CC remain unclear.Methods: In total, 298 patients with CTO who underwent retrograde septal CC PCI between January 2015 and May 2019 were retrospectively analyzed. Clinical data were compared to investigate the predictors of initial microcatheter tracking failure.Results: The initial and final microcatheter tracking success rates were 79.2% (236/298) and 96.6% (288/298), respectively. The procedural success rate was 94.0% (280/298). The right coronary artery-to-left anterior descending artery septal ratio (48.4% vs 33.1%, p=0.037) and CC tortuosity (34.6% vs 20.8%, p=0.045) were significantly higher in the initial microcatheter CC tracking failure group than in the successful tracking group. Multivariate logistic regression analysis revealed that severe collateral tortuosity (odds ratio [OR]: 13.241, 95% confidence interval [CI]: 3.429– 27.057, p=0.038), CC entry angle of < 90° (OR:4.921, 95% CI: 1.128– 9.997, p=0.002), CC exit angle of < 90° (OR:5.037, 95% CI: 2.237– 11.182, p=0.004), use of Finecross MG as initial microcatheter (OR:1.826, 95% CI: 1.127– 3.067, p=0.035), and shunning initial retrograde application of Guidezilla (OR:0.321, 95% CI: 0.267– 0.915, p=0.024) were variables independently associated with initial microcatheter CC tracking failure in patients with CTO undergoing retrograde septal CC PCI.Conclusion: The overall initial microcatheter CC tracking failure was 20.8%. Severecollateral tortuosity, CC entry, and exit angle of < 90°, use of Finecross MG as initial microcatheter, and shunning initial retrograde application of Guidezilla were variables independently associated with initial microcatheter CC tracking failure in patients with CTO undergoing retrograde septal PCI.Keywords: coronary chronic total occlusion, retrograde microcatheter tracking, collateral channel, predictor

Objectives. To assess the effectiveness and safety of ARW for vascular recanalization in CTO patients. Background. Chronic total occlusion (CTO) of coronary artery accompanied with large branch distal to the occluded segment (

A large, crack-free ZnGeP2 single crystal with size of Φ26 mm×70 mm was grown in a vertical three-zone tubular furnace by modified vertical Bridgman method, i.e. real-time temperature compensation technique with small temperature gradient in double-wall quartz ampoule. The as-grown single crystal was characterized by X-ray diffractometer (XRD), energy dispersive spectrometer (EDS), Fourier transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS). It was found that there is a face of (100) and its second-order XRD peaks were observed. The vertical elements distribution of the main part of the grown crystal has a stoichiometric ratio which is close to the ideal stoichiometry of 1:1:2. The IR transmittance of a sample of 2.5 mm thickness is above 58% in the range from 3500 to 800 cm-1. All these results demonstrate that the quality of the ZnGeP2 single crystal grown by the modified method is good, and could be used in the preparation of devices.

Objective To evaluate the effect of Zhuanggu Jianxi Decoction (壮骨健膝方, ZGJXD) on interleukin-1 β (IL-1 β)-induced degeneration of chondrocytes (CDs) as well as the activation of caveolin-p38 mitogen-activated protein kinase (MAPK) signal pathway, investigating the possible molecular mechanism that ZGJXD treats osteoarthritis.

The characteristics of real-time, synchronization, flexibility and reliability make WorldFIP Fieldbus suitable for locomotive network communication. This article describes the design of locomotive network test platform based on WorldFIP both hardware and software perspective. World FIP station can make date communication by writing applications in LABVIEW environment which verifies the correctness and reliability of the test platform. The design of test platform plays an important role for in-depth understanding of foreign locomotive core technology and lays a solid foundation for accelerating the domestic heavy-haul locomotive and development of products with independent intellectual property rights.

Background: It has been reported that coronary slow flow (CSF) is associated with acute myocardial infarction, ventricular tachycardia, ventricular fibrillation, and even sudden cardiac death. Although studies concerning the etiopathogenesis of CSF are scarce, diffuse atherosclerosis and endothelial dysfunction are thought to play important roles. It has been suggested that a high plasma thrombomodulin (TM) level seems to play an important role in the pathogenesis of atherosclerosis and endothelial dysfunction. Objectives: We hypothesized that a high plasma TM level might be associated with CSF and aimed to research the relationship between plasma TM level and CSF. Methods: Fifty-two CSF patients with angiographically proven CSF and 44 cases with normal coronary flow were included in this study. Coronary flow velocity was determined by the thrombolysis in myocardial infarction (TIMI) frame count method. Plasma TM levels were measured in all the study subjects. Results: Plasma TM levels were significantly higher in the CSF group compared to the control group (3.9 ± 0.5 vs. 3.6 ± 0.3 ng/mL, p = 0.01). There was a positive relationship (r = 0.31, p = 0.002) between plasma TM level and mean TIMI frame count (TFC). Factors associated with mean TFC were plasma TM level (β = 0.206, p = 0.038) and red cell distribution width (β = 0.088, p = 0.009) in multiple linear regression analysis. Conclusions: Patients with CSF have a higher plasma TM level, and this may play an important role in the pathogenesis of CSF. An elevated plasma TM level may be a predictor of CSF. Future studies are needed to confirm these results.

Some persistent organic pollutants (POPs) have been found in human semen but until this point it was unclear whether polybrominated diphenyl ethers (PBDEs) could be detected in human semen. In this study, PBDEs were found for the first time in human semen samples (n=101) from Taizhou, China. The concentrations of total PBDEs (∑PBDEs) varied from 15.8 to 86.8 pg/g ww (median=31.3 pg/g ww) and 53.2 to 121 pg/g ww (median=72.3 pg/g ww) in semen and blood samples, respectively. The ∑PBDE level in semen was about two times lower than in human blood, which was different in the distribution in the two matrices from other POPs. A correlation of ∑PBDE concentration was found between paired semen and in blood. The results suggest that semen could be used to detect PBDE burden in human body as a non-invasive matrix. In addition, the levels of BDE-209 and BDE-153, especially the latter, were much higher in blood than in semen, while the levels of BDE-28, BDE-47 and BDE-99 were comparable in the two matrices, suggesting that low brominated congeners could be more easily transferred to semen than high brominated congeners. Considering different toxicities among the PBDE congeners, it might be more significant to measure PBDEs in semen than in blood for evaluating male reproduction risks of PBDEs. Keywords: Polybrominated diphenyl ethers, Human, Semen, Blood, Non-invasive matrix

To study adverse effects and underlying mechanisms of beta-cypermethrin (β-cyp) on male reproductive system, the 15-day intact male adult Sprague-Dawley (SD) rats assay was used as an in vivo test. Male adult SD rats were treated by oral gavage with 0, 15 and 30mgβ-cyp/kgBW for 15 days. After 15-day treatments, the testes, epididymis and seminal vesicles were excised and weighed, respectively. One testis was used for testicular sperm head counts, and the other was for immunohistochemistry test to characterize the expression of androgen receptors (ARs). There were substantial decreases of both sperm head counts and daily sperm production after β-cyp exposure. The expression of AR decreased significantly in rats treated with 15 and 30mgβ-cyp/kgBW, and the gray scale pixel values in the three groups (0, 15 and 30mgβ-cyp/kgBW) were 113.79±13.58, 96.09±5.95 and 77.27±5.44, respectively. These findings suggested β-cyp has significant adverse effects on the reproductive system. Reducing the expression of AR is a potential mechanism of decreased sperm production caused by β-cyp.

ObjectivesThe strategies of repair of tetralogy of Fallot change with the age of patients. In children older than 4 years and adults, the optimal strategy may be to use different method of reconstruction of the right ventricular outflow tract from those followed in younger children, so as to avoid, or reduce, the pulmonary insufficiency that is increasingly known to compromise right ventricular function.MethodsFrom April, 2001, through May, 2008, we undertook complete repair in 312 patients, 180 male and 132 female, with a mean age of 11.3 years ±0.4 years, and a range from 4 to 48 years, with typical clinical and morphological features of tetralogy of Fallot, including 42 patients with the ventriculo-arterial connection of double outlet right ventricle. The operation was performed under moderate hypothermia using blood cardioplegia. The ventricular septal defect was closed with a Dacron patch. When it was considered necessary to resect the musculature within the right ventricular outflow tract, or perform pulmonary valvotomy, we sought to preserve the function of the pulmonary valve by protecting as far as possible the native leaflets, or creating a folded monocusp of autologous pericardium.ResultsThe repair was achieved completely through right atrium in 192, through the right ventricular outflow tract in 83, and through the right atrium, the outflow tract, and the pulmonary trunk in 36 patients. A transjunctional patch was inserted in 169 patients, non-valved in all but 9. There were no differences regarding the periods of aortic cross-clamping or cardiopulmonary bypass. Of the patients, 5 died (1.6%), with no influence noted for the transjunctional patch. Of those having a non-valved patch inserted, three-tenths had pulmonary regurgitation of various degree, while those having a valved patch had minimal pulmonary insufficiency and good right ventricular function postoperatively, this being maintained after follow-up of 8 to 24-months.ConclusionsBased on our experience, we suggest that the current strategy of repair of tetralogy of Fallot in older children and adults should be based on minimizing the insertion of transjunctional patches, this being indicated only in those with very small ventriculo-pulmonary junctions. If such a patch is necessary, then steps should be taken to preserve the function of the pulmonary valve.

Tumor-specific cytotoxic T lymphocytes (CTLs) from patients with early-stage tumors are usually more efficient at attacking tumor cells than CTLs from the progressing tumor stages. The authors investigated the antitumor activity of CTLs from gastric cancer patents and healthy donors. In this study, peripheral blood lymphocytes (PBLs) from gastric cancer patients and healthy donors were stimulated with HLA-A matched allogeneic gastric cancer cells such as KATO-3, MKN45, and SGC7901.Three different populations of lymphocyte, p5-CTL-KATO, h4-CTL-MKN45, and h4-CTL-KATO, were induced and expanded. Flow cytometry analyses showed that 85.2% to 97.8% of these cells were CD3-positive and 45.5% to 51.2% were CD8 - positive. The induced CTLs efficiently kill HLA-A2 or HLA-A24 gastric cancer cells through CTL-mediated cytotoxicity. However, no effects were observed for other cancer cells or HLA-A2 negative gastric cancer cells. The specific cytotoxicity of the induced CTLs was further confirmed by cold-target inhibition and monoclonal antibody blockage. These results suggest that CTL-mediated cytotoxicity specific for tumor cells can be produced by stimulating PBLs from healthy donors using HLA-A matched tumor cells, which will lead to the development of new immunotherapeutic strategies to kill cancer cells.

To observe the effect of naringin of Drynaria Rhizome, a Chinese medical component of Zhuanggu Jianxi Recipe (ZJR) containing serum on caveolin-p38MAPK signal factors (such as caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α) in IL-1β induced rabbit degenerated chondrocytes, and further to explore its mechanism for protecting articular cartilages.Naringin of Drynaria Rhizome was obtained and analyzed by HPLC-TOF/MS. Four weeks old New Zealand rabbits were killed and their bilateral knee joints were isolated aseptically. CDs were isolated and then cultured in vitro. The second generation of CDs were used for later experiment. The effect of naringin on CDs proliferation was detected by MTT assay. The effect of naringin on the expression of IL-1β-induced collagen II in CDs was detected by immunohistochemical method. The effect of naringin on caveolin-1, p-p38, and p-ATF-2 protein in IL-1β-induced CDs was detected by Western blot. The effect of naringin on mRNA expression of IL-1β and TNF-α in IL-1β-induced CDs was detected by RT-PCR.The appearance time of naringin in flow graphs of naringin standard solution and ZJR containing serum was 23.5 min, and the molecular weight ranged between 581.0 and 581.5 m/z. Naringin could promote the proliferation of CDs, and inhibit the effect of IL-1β on collagen II in CDs. Compared with the model group, naringin could reduce the expression of caveolin-1, p-p38, p-ATF-2, IL-1β, and TNF-α in IL-1β induced CDs (P0.05), which was approximate to the level of the normal group.Naringin could not only promote the proliferation of CDs, but also protect IL-1β-induced CDs. Its mechanism might be associated with decreasing the expression of caveolin-1, p-p38, and p-ATF-2 proteins, inhibiting caveolin-p38MAPK signal pathway, and further reducing mRNA expression of IL-1β and TNF-α in the downstream of caveolin-p38MAPK signal pathway.

Background It is becoming increasingly necessary for community health centers to make rehabilitation services available to patients with osteoarthritis of the knee. However, for a number of reasons, including a lack of expertise, the small size of community health centers and the availability of only simple medical equipment, conventional rehabilitation therapy has not been widely used in China. Consequently, most patients with knee osteoarthritis seek treatment in high-grade hospitals. However, many patients cannot manage the techniques that they were taught in the hospital. Methods such as acupuncture, tuina, Chinese medical herb fumigation-washing and t’ai chi are easy to do and have been reported to have curative effects in those with knee osteoarthritis. To date, there have been no randomized controlled trials validating comprehensive traditional Chinese medicine for the rehabilitation of knee osteoarthritis in a community health center. Furthermore, there is no standard rehabilitation protocol using traditional Chinese medicine for knee osteoarthritis. The aim of the current study is to develop a comprehensive rehabilitation protocol using traditional Chinese medicine for the management of knee osteoarthritis in a community health center. Method/design This will be a randomized controlled clinical trial with blinded assessment. There will be a 4-week intervention utilizing rehabilitation protocols from traditional Chinese medicine and conventional therapy. Follow-up will be conducted for a period of 12 weeks. A total of 722 participants with knee osteoarthritis will be recruited. Participants will be randomly divided into two groups: experimental and control. Primary outcomes will include range of motion, girth measurement, the visual analogue scale, and results from the manual muscle, six-minute walking and stair-climbing tests. Secondary outcomes will include average daily consumption of pain medication, ability to perform daily tasks and health-related quality-of-life assessments. Other outcomes will include rate of adverse events and economic effects. Relative cost-effectiveness will be determined from health service usage and outcome data. Discussion The primary aim of this trial is to develop a standard protocol for traditional Chinese medicine, which can be adopted by community health centers in China and worldwide, for the rehabilitation of patients with knee osteoarthritis. Trial registration Clinical Trials Registration: ChiCTR-TRC-12002538

From March to May, 2010, a pot experiment was conducted to investigate the effects of Eucalyptus grandis leaf litter at its early stage of decomposition on the growth and photosynthetic characteristics of Cichorium intybus. Four treatments with different application rate of the leaf litter, i.e., 0 g x pot(-1) (CK), 30 g x pot(-1) (A1), 60 g x pot(-1) (A2), and 90 g x pot(-1) (A3), were installed. Each pot contained 12 kg soil mixed with the leaf litter, and then, C. intybus was sown. The growth indicators of the C. intybus were measured at the 30, 45, 60, and 75 d after sowing, and the photosynthetic characteristics of the C. intybus in treatment A3 were studied after the seedlings third leaf fully expanded. At each measured time, the biomass accumulation and leaf area growth of C. intybus in treatments A1, A2, and A3 were inhibited significantly. At the early stage of the leaf litter decomposition, the synthesis of photosynthetic pigments of the C. intybus seedlings was inhibited significantly, and the inhibition effect was getting stronger with the increasing amount of the leaf litter addition. The diurnal change of the seedlings photosynthetic rate in all treatments showed a bimodal curve with midday depression, the stomatal conductance and water use efficiency had the same variation trend with the net photosynthetic rate, and the total diurnal photosynthesis decreased in the order of CKA1A2A3. The GC-MS analysis showed there were 33 kinds of small molecule compounds released gradually with the decomposition of the leaf litter, among which, allelopathic substance terpenoid dominated.

To analyze early mortality risk factors and clinical characteristics in our patients undergoing coronary artery bypass graft (CABG) surgery.Clinical data of 310 consecutive patients undergoing CABG from January 2005 to March 2007 were collected. Twenty-two risk factors were evaluated by univariate and multivariate Logistic stepwise regression analysis.Univariate statistical analysis revealed that factors significantly correlated with early death were 12 variables including age, diabetes, neurological dysfunction, old myocardial infarction, acute myocardial infarction, ejection function, left main artery stenosis, emergency procedure, cardiopulmonary bypass time, aortic cross-calming time, and mechanical ventilation time. Logistic stepwise regression analysis showed that emergency procedure, ejection function, age, cardiopulmonary bypass time, and mechanical ventilation time were independent risk factor of early mortality after procedure.Emergency procedure, ejection function, age, cardiopulmonary bypass time, and mechanical ventilation time are independent risk factors of early mortality after CABG procedure.

To estimate the clinical effect and long-term follow-up of minimally invasive direct coronary artery bypass grafting (MIDCAB) via left anterior small thoracotomy.MIDCAB via left anterior small thoracotomy approach was performed in 38 cases of coronary artery disease with single or multi-vessel involvement from January 2002 to October 2006. There were 25 males and 13 females with a mean age of (63.3 +/- 11.1) years old. The left internal mammary artery (LIMA) was harvested under direct vision or with the assistance of thoracoscopy. After heparinization the pericardium was directly opened to expose the target vessels. The coronary artery bypass grafting was completed on beating heart.The procedure were smoothly completed in all the 38 cases. The LIMA was anastomosed to the left anterior descending artery (LAD) or the diagonal artery in 20 cases, while two bypass grafts were performed in 8 cases (including 3 cases of sequential grafting and 5 cases of LIMA-radial artery Y-shaped grafting). Hybrid procedure was performed on 10 patients. There were no serious postoperative complications and operative deaths found. All the 38 cases except one were followed for 26 to 82 (53.2 +/- 28.5) months and no myocardial infarction or death occurred. NYHA class were I in 26 cases and II in 12 cases. There were 3 recurrence of angina, 2 patients relieved their symptoms with medication while 1 patient received stent implantation because of anastomose stenosis confirmed by coronary angiography 2 years after surgery.MIDCAB via left anterior small thoracotomy has low rate of mortality and adverse cardiac events. The long-term follow-up is good.

To verify the predictive value of EuroSCORE of early mortality in coronary artery bypass grafting (CABG) patients.From January 2005 to March 2007, 310 consecutive patients were operated with CABG. Detailed data for the EuroSCORE risk factor were collected and all patients were scored according to the EuroSCORE additive model, retrospectively or prospectively. Expected or predicted mortality was calculated for individual patients using the EuroSCORE algorithms, arranged sequentially in order of predicted score. The population was divided into three clinically relevant risk categories according to the range of predicted mortality rate. Expected mortality was compared to observed or actual mortality for each risk category. Mortality was defined as death from any cause within 30 days of operation or within the same hospital admission.Preoperative overall patients: low-risk group was 25.2% (78/310), middle-risk group was 48.4% (150/310), high-risk group was 26.4% (82/310). In the EuroSCORE model, predicted mortality was 1.4% for low-risk group, 2.7% for middle-risk group, 7.4% for high-risk group, and 3.6% for overall patients. Actual mortality was 0, 1.3% and 3.7% respectively, overall early mortality was 1.6%. Area under the ROC curve was 0.78.The EuroSCORE yield good predictive value for hospital mortality of patients undergoing CABG, especially in off-pump CABG.

MG7-Ag is a human gastric-carcinoma-associated antigen with a high specificity. So far it is remained unclear whether MG7-Ag is correlated with the in vivo cellular immune response of patients with gastric cancer. In this study, we detected the expression of the T cell receptor (TCR) repertoire of T cell subpopulations and cytokines in tumor-infiltrating lymphocytes (TIL), peripheral blood lymphocytes (PBL), and residue benign mucosal lymphocytes (NML) of patients with gastric cancer using semiquantitative RT-PCR. Our data showed that the expanded clones in CD8+NML and TIL and CD4+NML and PBL in MG7-Ag-positive patients were significantly fewer than those of MG7-Ag-negative patients (p = 0.0360; p = 0.0026; p = 0.0065 p = 0.0109, respectively). The levels of IL-8 in CD8+TIL and TNF in CD4+TIL from the MG7-Ag-positive group were significantly higher than those from the MG7-Ag-negative group (p = 0.0302; p = 0.0177, respectively). Taken together, the results demonstrated a weaker T cell immune response and more proinflammatory cytokine secretion in MG7-Ag-positive patients with gastric cancer than in MG7-Ag-negative ones. This likely contributes to the poor prognosis in MG7-Ag-positive gastric-cancer patients.Key words: gastric carcinoma, tumor-associated antigen, T cell receptor repertoire, cytokine, RT-PCR.

To screen bioactive peptides that mimic the epitope of gastric cancer associated antigen.Anti-gastric cancer monoclonal antibody (mAb) MG7 was purified by ion chromatography, and then coated on ELISA plate. By using the mAb as selective molecular, a 12-meres phage displaying peptide library was biopanned, and the positive clones were selected.12 positive phage clones were selected.A candidate mimic epitope of gastric cancer associated antigen was identified, which provided the foundation for developing tumor peptide vaccine.

To study the relationship between blood pressure control status and patients' knowledge on hypertension prevention and control among hypertensive patients.A total of 726 hypertensives were selected from four community health service centers (2 urban and 2 rural) in Beijing. All subjects were investigated by questionnaires and their blood pressures were measured at the same time.The rate for blood pressure under control (140/90 mm Hg, 1 mm Hg = 0.133 kPa) in the rural and urban patients were 46.4% and 23.9% respectively. The control rate increased with the increase of patients' knowledge on prevention and control of hypertension in both urban and rural patients. The cumulative effect of knowledge on hypertension control status could contribute 30.0% to the difference in hypertension control rate between rural and urban patients.Patients' knowledge on hypertension control was significantly related to the rate on hypertension control. Health education should be helpful to improve the rate on hypertension control.

To investigate the overexpression of prion protein (PrP) in drug-resistant gastric cancer cell line SGC7901/ADR and its role in multidrug resistance in gastric cancer.(1) The expression of PrP in SGC7901/ADR, SGC790/VCR and their parental cell line SGC7901 was detected with Northern and Western blot at the mRNA and protein level. (2) Eukaryotic sense and antisense expression vector were constructed based on DNA recombination technology and (3) introduced into SGC7901 and SGC7901/ADR cell lines through electroporation. (4) The accumulation and retention of ADR in transiently transfected cells were detected by flow cytometry.(1) Northern and western blot suggested significantly higher expression of PrP in SGC7901/ADR and SGC7901/VCR than that in SGC7901. (2) 48 hours after the vectors transfection, the average fluorescence intensity of Adr in transfected cells were detected. The accumulation intensity were 8.9 +/- 0.7 in BS, 6.6 +/- 0.3 in PS and 7.5 +/- 0.6 in PA. The retention intensity were 9.3 +/- 0.6 in SGC7901, 5.9 +/- 0.5 in PS and 7.1 +/- 0.5 in PA. There were significant difference between PS and BS with P0.01, as well as RA and BA with P0.01. These data suggested that PrP gene could affect the drug accumulation in gastric cancer cells after its transfected into cells.PrP was highly expressed in gastric cancer cell lines SGC7901/ADR and SGC7901/VCR. Overexpression of PrP had certain effect on drug accumulation in gastric cancer cells.

To investigate the differential expression of RPL6/Taxreb107 between drug-resistant gastric cancer cell line SGC7901/ADR and gastric cancer cell line SGC7901 as well as its correlation with multiple-drug resistance (MDR) in gastric cancer cells.Total RNA was extracted from SGC7901 and SGC77901/ADR, with internal control RT-PCR, Northern blot, gene cloning and expression, construction of eukaryotic expression vector, gene transfection by electroporation. The accumulation and retention of ADR in transiently transfected cell was detected by flow cytometry.The internal control RT-PCR and Northern blot showed high RPL6/Taxreb107 expression in SGC7901/ADR cell line. Sense and antisense eukaryonic expression vectors demonstrated by double enzyme digestion were successfully transfected into gastric cancer cell line SGC7901 and SGC7901/ADR respectively by electroporation. The accumulation and retention of ADR detected 48 hours after transfection showed that RPL6 gene had shown effect on drug resistance in gastric cancer cell.The high expression of RPL6/Taxreb107 in drug resistant gastric cancer cell shows its correlation with multiple-drug resistance in gastric cancer.

MC5 is a murine monoclonal antibody with a good specificity to human colorectal carcinoma and smaller murine antibody can significantly decrease the possibility of developing human antimouse antibody response in vivo study. The aim of this study was to prepare single chain variable fragments (ScFv) of MC5.mRNA was isolated from the hybridoma cell line producing MC5, and the DNAs encoding variable domains of heavy and light chains(VH and VL DNAs) of the antibody were amplified separately by RT-PCR and assembled into ScFv DNAs with a linker DNA. The ScFv DNAs were ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into E. coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage antibody ScFvs. After two rounds of panning with cell line SW480 highly expressing MC5-binding antigen, the phage clones displayed ScFv fragments of the antibody were selected by ELISA, and the affinity of the positive phage clones was assayed by competition ELISA.The VH, VL, and ScFv DNAs were about 340 bp, 320 bp, and 750 bp, respectively. Ten phage clones displayed ScFv of MC5 were selected from 25 enriched phage clones, and 3 of the 10 phage clones had higher affinity of binding to the antigen.The phage-displayed ScFv fragments of monoclonal antibody MC5 are successfully produced by phage display technique, which may provide a way for broadening the application range of the antibody.

To generate soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal and gastric carcinomas.mRNA was isolated from the hybridoma cell line producing MC3 and the DNAs encoding variable domains of heavy and light chains (VH and VL) of the antibody were amplified separately by RT-PCR and assembled into ScFv DNA with a linker DNA. The ScFv DNA was ligated into the phagemid vector pCANTAB5E and the ligated sample was transformed into E.coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phages. After two rounds of panning with gastric carcinoma cell line AGS highly expressing MC3-binding antigen, the phage clones displaying ScFv fragments of the antibody were selected by ELISA. 4 phage clones showing strong signal in ELISA were used to infect E.coli HB2151 to express soluble ScFvs. The soluble ScFvs were identified by Dot blot and Western blot, and their antigen-binding activity was assayed by ELISA. The VH and VL DNAs of the ScFv DNA derived from phage clone 19 were sequenced.The VH,VL and ScFv DNAs were about 340 bp, 320 bp and 750 bp respectively. After two rounds of panning to the recombinant phages, 18 antigen-positive phage clones were selected from 30 preselected phage clones by ELISA. All the soluble ScFvs derived from the 4 out of the 18 antigen-positive phage clones were about M(r)32000 and concentrated in periplasmatic space under the given culture condition. The soluble ScFvs could bind the antigen, and they shared the same binding site with MC3. The sequences of the VH and VL DNAs of the MC3 ScFv showed that the variable antibody genes belonged to the IgG1 subgroup,kappa-type.The soluble ScFv of MC3 is successfully produced, which not only provides a possible novel targeting vehicle for in vivo and in vitro study on associated cancers, but also offers the antibody a stable genetic source.

AIM: To evaluate the immunity of chemically modified tumor cell vaccine. METHODS: Tumor cell vaccines (TCV) were prepared by incubating the live Ehrlich ascites tumor cells with concanavalin A-mitomycin C (ConA-MMC), mitomycin C (MMC), concanavalin A-glutaraldehyde (ConA-Glu), glutaraldehyde ( Glu ), or paraformaldehyde ( Para ), respectively. The whole cell or soluble forms of the vaccines were administered intraperitoneally into Kunming mice once a week for three times prior to the intraperitoneal inoculation of a lethal dose of live tumor cells. A second challenge with live tumor cells was given four weeks later. Survival and antibody production of the mice were analyzed. RESULTS: After the first challenge, the mice, received whole TCV of ConA-MMC, MMC (P < 0.01) and Glu (P < 0.05) promoted survival incidence than the controls. All the treated mice had the survival time prolonged. ConA-MMC vaccine treated mice had longer survival days than that of ConA-Glu ones (P < 0.05). For the soluble TCV immunized mice, those treated with vaccines of Para (P < 0.01), ConA-Para and ConA-Glu (P < 0.05) had longer survival periods compared with that of the controls. Following the second challenge, survival incidence of the mice received vaccines of ConA-MMC, MMC, ConA-Glu or Glu was significantly increased (P < 0.01). Moreover, all the treated mice had the survival time prolonged, and ConA-MMC vaccine treated mice had longer survival days than that of Para treated ones (P < 0.05). Antibodies against Ehrlich ascites tumor cells were found to be positive in sera of the mice treated with whole TCV of ConA-MMC. CONCLUSION: Ehrlich ascites tumor cells are immunogenic when treated with ConA-MMC, MMC, ConA-Glu, Glu or Para, which might act as safe and effective tumor vaccines with safety and effectiveness.

When clock frequencies exceed gigahertz, the skin depth in analog and digital circuits greatly decreases. The irregular shape of the cross section of the interconnect plays an increasingly important role in interconnect parasitic extraction. However, existing methods only focus on the rough surface of the interconnect, while ignoring other irregular shapes, such as the trapezoidal cross section. In this work, a new simulation method is proposed for irregular interconnects, which is applicable to arbitrary irregular shapes and to a wide range of frequencies. The method involves generating a mesh information file firstly and then extracting the frequency-dependent resistance based on a numerical solution of scalar wave modeling by using the method of moments. The singularity extraction method is used to calculate the self-inductors. The data from experiments verify the accuracy of our proposed method.

AIM:To study the feasibility of panning and screening phage-displaying recombinant single-chain variable fragment (ScFv) of anti-tumor monoclonal antibodies for fixed whole cells as the carriers of mAb-binding antigens. METHODS: The recombinant phage displaying libraries for anti-colorectal tumor mAb MC3Ab, MC5Ab and anti-gastric tumor mAb MGD1 was constructed. Panning and screening were carried out by means of modified fixation of colorectal and gastric tumor cells expressed the mAb-binding antigens. Concordance of binding specificity to tumor cells between phage clones and parent antibodies was analyzed. The phage of positive clones was identified with competitive ELISA, and infected by E.coli HB2151 to express soluble ScFv. RESULTS:The ratio of positive clones to MC3-ScF-MC5ScFv and MGD1-ScFv were 60 %, 24 % and 30 %. MC3ScFv had Mr 32 000 confirmed by Western blot. The specificity to antigen had no difference between 4 positive recombinant phage antibodies and MC3Ab. CONCLUSION:The modified process of fixing whole tumor cells is efficient, convenient and feasible to pan and screen the phage–displaying ScFv of anti-tumor monoclonal antibodies.

Novel cDNA fragment associated with gastric cancer drug resistance was screened out from a library by monoclonal antibody MGr1