Correction to "Gut microbiota dysbiosis in Chinese children with type 1 diabetes mellitus: An observational study".

[This corrects the article on p. 2394 in vol. 27, PMID: 34040330.].

Effects of 2,2',4'-trihydroxychalcone on the proliferation, metastasis and apoptosis of A549 human lung cancer cells.

The aim of the present study was to evaluate the antitumor effects of 2,2',4'-trihydroxychalcone (7a) on the A549 human lung cancer cell line. A549 cells were treated with different concentrations of 7a for different time periods. Cells without 7a were used as the negative control group. Cell proliferation, invasion, vasculogenic mimicry (VM) formation, heterogeneous adhesion and apoptosis were measured using Cell Counting Kit-8, Transwell invasion, VM, adhesion and flow cytometric assays, respectively. In addition, the expression of related proteins was determined using western blot analysis or ELISA. The present study found that 7a had a significant inhibitory effect on the survival rate of the A549 lung cancer cells but almost no effect on BEAS-2B human lung epithelial cells or human venous endothelial cells. The migration rate, VM length, invasion rate and heterogeneous adhesion number of cells treated with 7a significantly decreased as the concentration increased, while the apoptosis rate increased. Western blot analysis showed that 7a treatment significantly increased the expression levels of E-cadherin, cleaved poly (ADP-ribose) polymerase, Bax and caspase-3 and simultaneously decreased the expression levels of metalloproteinase-2/9, Bcl-2, phosphorylated (p)-PI3K, p-AKT, p-mTOR, vascular endothelial growth factor (VEGF), E-selectin and N-cadherin. At the same time, the ELISA results showed that the level of the pro-angiogenic factor VEGF in the culture media was reduced in the presence of 7a. In addition, 7a could also reduce the nuclear NF-κB protein expression, which could inhibit the gene transcription of tumor apoptosis and metastasis-related proteins. Therefore, 7a may exert inhibitory effects on A549 cells by inhibiting cell proliferation, migration, VM formation and heterogeneous adhesion, as well as by inducing apoptosis through the suppression of the PI3K/AKT/NF-κB signaling pathway; these findings suggested that 7a may be a promising agent for the treatment of lung cancer.

Gut microbiota dysbiosis in Chinese children with type 1 diabetes mellitus: An observational study.

BACKGROUND: Gut microbiota dysbiosis is reportedly actively involved in autoimmune diseases such as type 1 diabetes mellitus (T1DM). However, the alterations in the gut microbiota and their correlation with fasting blood glucose (FBG) in Chinese children with T1DM remain unclear. AIM: To investigate alterations in the gut microbiota in Chinese children with T1DM and their associations with clinical indicators. METHODS: Samples from 51 children with T1DM and 47 age-matched and gender-matched healthy controls were obtained, to explore the structural and functional alterations in the fecal microbiota. The V3-V4 regions of the 16S rRNA gene were sequenced on a MiSeq instrument, and the association with FBG were analyzed. RESULTS: We found that the bacterial diversity was significantly increased in the T1DM-associated fecal microbiota, and changes in the microbial composition were observed at different taxonomic levels. The T1DM-reduced differential taxa, such as Bacteroides vulgatus ATCC8482, Bacteroides ovatus, Bacteroides xylanisolvens, and Flavonifractor plautii, were negatively correlated with FBG, while the T1DM-enriched taxa, such as Blautia, Eubacterium hallii group, Anaerostipes hadrus, and Dorea longicatena, were positively correlated with FBG. Bacteroides vulgatus ATCC8482, Bacteroides ovatus, the Eubacterium hallii group, and Anaerostipes hadrus, either alone or in combination, could be used as noninvasive diagnostic biomarkers to discriminate children with T1DM from healthy controls. In addition, the functional changes in the T1DM-associated fecal microbiota also suggest that these fecal microbes were associated with altered functions and metabolic activities, such as glycan biosynthesis and metabolism and lipid metabolism, which might play vital roles in the pathogenesis and development of T1DM. CONCLUSION: Our present comprehensive investigation of the T1DM-associated fecal microbiota provides novel insights into the pathogenesis of the disease and sheds light on the diagnosis and treatment of T1DM.

Solution and rheological properties of cationic cellulose/gemini surfactant: Effect of the alkyl chain and spacer length.

In this paper, three sulfonate-containing gemini surfactants, sodium 1,1'-(4,4'-methylenebis(4,1-phenylene))bis(1-oxooctane-2-sulfonate) (C8-M1-C8), sodium 1,1'-(4,4'-(ethane-1,2-diyl)bis(4,1-phenylene))bis(1-oxooctane-2-sulfonate) (C8-M2-C8), sodium 1,1'-(4,4'-methylenebis(4,1-phenylene))-bis(1-oxododecane-2-sulfonate) (C12-M2-C12), were synthesized and characterized with FT-IR, 1H NMR and MS. Furthermore, interaction between a cationic cellulose-based polyelectrolyte, PQ-10, and gemini surfactants were investigated by surface tension, turbidity, flow and low-amplitude oscillation rheology analysis. For comparing, the interaction of their corresponding monomeric counterpart sodium dodecyl sulfate (SDS), sodium 1-octanesulfate (SOS) was also studied. Results showed that the concentration value at T1, defined as critical surface complex concentration, for the PQ-10/surfactant was in order of PQ-10/C8-M2-C8> PQ-10/C8-M1-C8 > PQ-10/C12-M2-C12. Precipitation appeared at low concentration for Gemini surfactants than their monomeric counterparts, and for the gemini surfactants with shorter spacer or longer hydrocarbon chain. The increase/decrease of the crossover frequency (ωc) (the relaxation time, τc) for PQ-10/C12-M2-C12 indicated the formation/collapse of network structures, while PQ-10/SDS showed no obvious change.

SOX2-induced upregulation of lncRNA LINC01561 promotes non-small-cell lung carcinoma progression by sponging miR-760 to modulate SHCBP1 expression.

Long noncoding RNAs (lncRNAs) have been shown to have critical regulatory roles in tumorigenesis. lncRNA LINC01561 (LINC01561) is a newly identified tumor-related lncRNA and its dysregulation has been demonstrated in several tumors. However, whether LINC01561 is involved in the progression of non-small-cell lung carcinoma (NSCLC) and its underlying mechanisms remain unknown. In this study, we first provided evidence that LINC01561 expressions were distinctly upregulated in NSCLC tissues and cell lines. Combining with bioinformatics assays and mechanism experiments, our group demonstrated that LINC01561 was activated by SOX2 in NSCLC. Clinical research revealed that upregulation of LINC01561 was related to poorer clinicopathologic features and shorter survival time. Functionally, suppression of LINC01561 exhibited tumor-suppressive functions through impairing cell proliferation, migration, and invasion as well as inducing apoptosis. Moreover, we verified that LINC01561 could directly bind to miR-760, isolating miR-760 from its target gene SHC SH2 domain-binding protein 1 (SHCBP1). We also found that SHCBP1 was lowly expressed in NSCLC and served as a tumor promoter. A functional study indicated that LINC01561 regulated SHCBP1 expression by competitively binding to miR-760. In summary, our findings indicated that SOX2-induced overexpression of LINC01561 promoted the proliferation and metastasis by acting as a competing endogenous RNA to modulate SHCBP1 by sponging miR-760.

Aspirin alleviates cardiac fibrosis in mice by inhibiting autophagy.

Aspirin (ASA) is a cardioprotective drug with anti-cardiac fibrosis action in vivo. This study was aimed to clarify the anti-cardiac fibrosis action of ASA and the underlying mechanisms. Two heart injury models (injection of isoproterenol and ligation of the left anterior descending branch) were used in mice to induce cardiac fibrosis. The animals were treated with ASA (10 mg·kg-1·d-1, ig) for 21 and 14 d, respectively. ASA administration significantly improved cardiac function, and ameliorated heart damage and fibrosis in the mice. The mechanisms underlying ASA's anti-fibrotic effect were further analyzed in neonatal cardiac fibroblasts (CFs) exposed to hypoxia in vitro. ASA (0.5-5 mmol/L) dose-dependently inhibited the proliferation and Akt phosphorylation in the CFs. In addition, ASA significantly inhibited CF apoptosis, and decreased the levels of apoptosis markers (cleaved caspase 3 and Parp1), which might serve as a side effect of anti-fibrotic effect of ASA. Furthermore, ASA dose-dependently inhibited the autophagy in the CFs, as evidenced by the reduced levels of autophagy marker LC3-II. The autophagy inhibitor Pepstatin A (PepA) promoted the inhibitory effect of ASA on CF proliferation, whereas the autophagy inducer rapamycin rescued ASA-caused inhibition of CF proliferation, suggesting an autophagy-dependent anti-proliferative effect of ASA. Both p38 inhibitor SB203580 and ROS scavenger N-acetyl-cysteine (NAC) significantly decreased Akt phosphorylation in CFs in the basal and hypoxic situations, but they both significantly increased LC3-II levels in the CFs. Our results suggest that an autophagy- and p38/ROS-dependent pathway mediates the anti-cardiac fibrosis effect of ASA in CFs. As PepA and SB203580 did not affect ASA-caused inhibition of CF apoptosis, the drug combination will enhance ASA's therapeutic effects.

Effects of different fatty acids composition of phosphatidylcholine on brain function of dementia mice induced by scopolamine.

BACKGROUND: Phosphatidylcholine (PC), the major source of dietary choline, has been demonstrated to improve the capability of learning and memory in rodent and the amelioration of long-chain n-3 polyunsaturated fatty acids (PUFA) on anti-aging and anti-oxidation is widely known as well. In this study, three kinds of PC were chose to demonstrate the role of different fatty acids composition on glycerol backbone in improving the brain function of mice induced by scopolamine which was used to impair cholinergic system and cause oxidative stress. METHODS: Male BALB/c mice were randomly divided into 5 groups: model (M) group, control (Con) group, egg yolk lecithin (EL) group, squid PC (SQ-PC) group and sea cucumber PC (SC-PC) group. The intraperitoneal injection of scopolamine hydrobromide (5 mg/kg) was carried out on the 8(th) of group feeding and sustained daily until the end of test. Morris water maze test was used to evaluate the improvement of cognitive decline and the activity of acetylcholinesterase (AchE), superoxide dismutase (SOD) and monoamine oxidase (MAO) and malondialdehyde (MDA) content in brain were measured to assess the physiological changes. RESULTS: In behavior test, the latency of PC groups was significantly reduced, while number of crossing the platform and time in target quadrant were increased in comparison with M group and the improvements of SQ-PC and SC-PC were better than that of EL (P < 0.05). Similar trend was observed in physiological changes. The AchE activity was effectively decreased and the SOD activity increased in hippocampus, cortex and white matter when comparing PC groups with M group. SQ-PC, SC-PC and EL respectively showed 22.82, 28.80 and 11.81 % decrease in MDA level in brain compared with M group. The MAO activity in white matter of SQ-PC, SC-PC and EL group separately depressed 33.05, 33.64 and 19.73 % in comparison with M group. No significance between SQ-PC and SC-PC was found in these indicators except the SOD activity in hippocampus and white matter. SQ-PC group had a higher SOD activity in hippocampus (103.68U/mg · prot.) and lower in white matter (120.57 U/mg · prot.) than SC-PC group (95.53 U/mg · prot. in hippocampus, 134.49 U/mg · prot. in white matter). PC rich in n-3 PUFA acted more ameliorative effects than that barely contained on the indicators above. CONCLUSIONS: Different fatty acids composition of PC all could diminish the cognitive decline and biological damage and protect the brain. EPA and DHA partly enhaced to the advantageous effects.

Identification of differentially expressed genes in American cockroach ovaries and testes by suppression subtractive hybridization and the prediction of its miRNAs.

Studies on the cockroach have contributed to our understanding of several important developmental processes, especially those that can be easily studied in the embryo. However, our knowledge on late events such as gonad differentiation in the cockroach is still limited. The major aim of the present study was to identify sex-specific genes between adult female and male Periplaneta americana. Two cDNA libraries were constructed using the suppression subtractive hybridization method; a total of 433 and 599 unique sequences were obtained from the forward library and the reverse library, respectively, by cluster assembly, and sequence alignment of 1,032 expressed sequence tags. The analysis of the differentially expressed gene functions allowed these genes to be categorized into three groups: biological process, molecular function, and cellular component. The differentially expressed genes were suggested to be related to the development of the gonads of P. americana. Twelve differentially expressed genes were randomly selected and verified using relative quantitative real-time polymerase chain reaction (qRT-PCR). Meanwhile, by adopting a range of filtering criteria, we predicted two potential microRNA sequences for P. americana, pam-miR100-3p and pam-miR7. To confirm the expression of potential microRNAs (miRNAs) in American cockroach, a qRT-PCR approach was also employed. The data presented here offer the insights into the molecular foundation of sex differences in American cockroach, and the first report for the miRNAs in this species. In addition, the results can be used as a reference for unraveling candidate genes associated with the sex and reproduction of cockroaches.

[Effect of curcumin on the learning, memory and hippocampal Ca+/CaMK II level in senescence-accelerated mice].

OBJECTIVE: To explore effect of curcumin in different concentrations on learning and memory of senescence-accelerated mice (SAM) and their possible mechanisms. METHODS: Mice were randomly divided into six groups: the SAMR1 normal control group, the SAMP8 model control group, the SAMP8 + solvent (the peanut oil) control group, SAMP8 + low, middle and high dose curcumin groups. Mice were gastrogavage for 25 successive days. On the next day of ending the experiment, changes of learning and memory in mice of each group were observed by Morris water maze. The hippocampal [Ca2+] was determined. Expressions of hippocampal calmodulin-dependent protein kinase II (CaMK II) and Calmodulin (CaM) mRNA were detected using Western blot and reverse transcription polymerase chain reaction (RT-PCR) respectively. RESULTS: The latency to find the hidden platform was remarkably prolonged, the hippocampal [Ca2+]i was markedly increased, the expression of CaMK II in the hippocampal membrane and the level of hippocampal CaM mRNA were significantly reduced in the SAMP8-model control group (P < 0.05, P < 0.01). The latency to find the hidden platform was remarkably shortened in the SAMP8 + middle dose curcumin and the SAMP8 + high dose curcumin groups (P < 0.01). The hippocampal [Ca2+]i was markedly lowered, the expression of CaMK II in the hippocampal membrane and the level of hippocampal CaM mRNA obviously increased in the SAMP8 + low, middle and high dose curcumin groups (P < 0.05, P < 0.01). CONCLUSION: Curcumin could improve learning and memory Ca2+/capacities of SAM by lowering hippocampal [Ca2+] overload, increase the hippocampal CaM mRNA level and CaMK II expression in the hippocampal dose-dependently.

[Characterization of atherosclerotic plaque in patients with unstable angina pectoris and stable angina pectoris by optical coherence tomography].

OBJECTIVE: To compare the characterization of coronary atherosclerotic plaques in patients with unstable angina pectoris (UAP) and stable angina pectoris (SAP) by optical coherence tomography (OCT). METHODS: OCT was performed in 47 patients (23 UAP and 24 SAP) undergoing coronary angiography. Lipid-rich plaque (defined by > or = 2 quadrants of the cross-section area), thin cap fibroatheroma (TCFA), thickness of fibrous cap, plaque rupture, calcification and thrombus visualized by OCT were compared between UAP and SAP patients. RESULTS: OCT imaging was successfully in 44 out of 47 patients (22 UAP, 22 SAP). Proportion of lipid-rich plaques was similar between UAP and SAP groups [91% (20/22) vs. 73% (16/22), P = 0.741]. The minimum thickness of fibrous cap in the UAP group was significantly thinner than that in SAP group [(69.5 +/- 34.7) microm vs. (141.1 +/- 68.5) microm, P = 0.000] and the rate of fibrous cap erosion in the UAP group was significantly higher than that in the SAP group [59% (13/22) vs. 9% (2/22), P = 0.000]. Percents of TCFA [73% (16/22) vs. 14% (3/22), P = 0.000] and plaque rupture [50% (11/22) vs. 9% (2/22), P = 0.003] were significantly higher in UAP group compared those in SAP group. Incidence of thrombus and calcification were similar between two groups. CONCLUSIONS: OCT imaging can clearly define plaque characterization of coronary atherosclerosis. UAP patients have thinner fibrous cap, higher incidences of fibrous cap erosion, plaque rupture and TCFA compared patients with SAP.

[Influence of avian reovirus infection on the Bursa and immune-reactions in chickens].

The influences of avian reovirus (ARV) infection at 1 day of age on bursa development, antibody responses after vaccinations to avian influenza virus (AIV), Newcastle disease virus (NDV) and infectious bursal disease virus (IBDV), and pathogenecity of virulent IBDV (v-IBDV) were studied in chickens of SPF-origin. The results indicated that LY strain ARV infection in 1-day-old chickens caused atrophy of the Bursa and decreased lymphocyte numbers in the bursa, but it gave no significant negative effects on growth rates and antibody titers to AIV and NDV after vaccination. LY strain ARV infection decreased antibody titers to IBDV in B87 vaccinated birds but all vaccinated birds infected with ARV were still full protected from death or clinical syndromes after v-IBDV challenge. Although all B87-vaccinated birds were full protected from death after v-IBDV challenges, the antibody titers to AIV or NDV after vaccinations with inactivated vaccines were significantly lower in v-IBDV challenged birds than controls. Supprisingly, ARV infection at ages of 1-7 days could compromise the immune suppression induced by v-IBDV in B87-vaccinated birds as HI antibody titers to AIV and NDV in ARV-infected groups were significantly higher than chicknes with no ARV infection. A discussion was made on the interactions between ARV infection and vaccine IBDV or v-IBDV to explain such sophisticated phenomena in the bird experiments.

[Anatomical study on Jingming (BL 1)].

OBJECTIVE: To explore the anatomical structures, and depth and direction of needling at Jingming (BL 1), so as to provide anatomical basis for its clinical application. METHODS: Forty-eight adult orbital specimens were observed by dissection. RESULTS: When a needle was vertically inserted into Jingming (BL 1), the needle tip will past through the skin, subcutaneous tissue, medial palpebral ligament, medialis rectus and orbital adipose body. Above the body of the needle, there are ophthalmic artery, anterior ethmoidal artery and nasociliary nerve. The average distance between the skin at the punctured point and the anterior ethmoidal artery is (18.25 +/- 4.45) mm, with an angle of (12.5 +/- 5.5) degrees, and the average distance between the skin at the punctured point and the optic nerve tunnel frontal point is (43.37 +/- 7.84) mm. CONCLUSION: To avoid bleeding caused by injuring the anterior ethmoidal artery, acupuncture at Jingming (BL 1) should avoid deeply inserting needled back-upwards and upwards, and the needling depth should not exceed 30.36 mm to avoid injury of the optic nerve tunnel frontal point.

[Visualization of intimal proliferation of drug-eluting stent and bare mental stent by use of optical coherence tomography].

OBJECTIVE: To compare neointimal proliferation of drug-eluting stent (DES) with bare mental stent (BMS) by optical coherence tomography (OCT). METHODS: OCT images were obtained in 21 diseased coronary vessels with 23 stents in 19 patients with coronary artery disease at 5 - 93 months post DES or BMS stents. Twenty-two stents of all 23 stents were divided into three groups. Nine DES stents at 6 - 10 months post stenting were considered as group A, 8 BMS stents at 5 - 10 months post stenting as group B, and 5 BMS stents at 23 - 93 months post stenting as group C. OCT images were quantitatively analyzed to compare neointimal proliferation of three groups after stenting. RESULTS: All 21 vessels and 23 stents OCT images were successfully acquired. The maximal neointima, luminal loss in diameter and cross sectional area (CSA), and restenosis in diameter and CSA were significantly statistically different within three groups. The maximal intimal proliferations post stenting in group A were significantly lower than group B (0.20 mm +/- 0.13 mm vs 0.81 mm +/- 0.46 mm, P = 0.019) or group C (0.91 mm +/- 0.27 mm, P = 0.007), luminal loss of diameter in group A were significantly lower than group B (0.27 mm +/- 0.17 mm vs 1.12 mm +/- 0.79 mm, P = 0.009) or group C (1.20 mm +/- 0.31 mm, P = 0.013), restenosis rates in diameter in group A were significantly less than group B (8.90% +/- 4.47% vs 36.36% +/- 24.34%, P = 0.009) or group C (35.48 +/- 6.09, P = 0.017), luminal loss in CSA in group A were lower than group B (1.14 mm(2) +/- 0.9 mm(2) vs 3.96 mm(2) +/- 2.62 mm(2), P = 0.009) or group C (4.66 mm(2) +/- 1.66 mm(2), P = 0.006), and restenosis rates in CSA in group A were less than group B (15.43% +/- 7.89% vs 48.14% +/- 30.43%, P = 0.017) or group C (55.20% +/- 11.24%, P = 0.009). Almost all surfaces of 13 BMS stent struts were covered by significant neointimal coverage, surfaces of 10 DES struts were less significantly neointimal coverage, and some surfaces of DES struts were uncovered with neointima even at 29 months post stenting. CONCLUSION: OCT imaging can clearly visualize stent struts and neointimal formation of strut surfaces post DES or BMS stenting, and this new imaging modality will play important role in evaluating the efficacy of drug-eluting stent.

[Coronary artery atherosclerotic plaque and stent visualizations by optical coherence tomography].

OBJECTIVE: To evaluate coronary artery atherosclerotic plaque characteristics and changes post coronary stenting by optical coherence tomography (OCT). METHODS: OCT images were obtained in 22 diseased coronary vessels after coronary angiography or percutaneous coronary interventions (PCI) in 20 patients and in 23 stents [7 sirolimus-eluting stents (SES) follow up at 4-29 months post stenting and 8 bare mental stents (BMS) at 4-35 months post stenting, 8 stents immediately after PCI]. RESULTS: All 22 vessels and 23 stents OCT images were successfully acquired. Two thromboses, 8 fibrous, 9 lipid-rich and 3 calcium plaques as well as 3 plaque ruptures were visualized by OCT. No significant neointimal proliferation and restenosis were found in SES stents and some struts were not covered with neointima even at 29 months post stenting. Significant neointimal proliferation on surfaces of stent struts were visualized in all 8 BMS stents and restenosis was detected in 3 BMS stents. OCT images obtained immediately after PCI showed that 3 stents were well positioned, tissue prolapse between coronary stent struts occurred in 4 stents and stent dissociation with vessel wall could be seen in 1 stent. CONCLUSIONS: OCT imaging can clearly visualize different types of atherosclerotic plaques. By providing detailed information on plaque characteristics, this technique might help cardiologists in choosing suitable stents and guiding preventive therapy for patients with coronary heart disease.

Changes of learning, memory and levels of CaMKII, CaM mRNA, CREB mRNA in the hippocampus of chronic multiple-stressed rats.

BACKGROUND: The effect of chronic stress on cognitive functions has been one of the hot topic in neuroscience. But there has been much controversy over its mechanism. Such single stressor applied in the past could not simulate complicated living circumstances that people confronted with. The aim of this study was to investigate the effects of chronic multiple-stress on learning and memory as well as on the levels of calcium/calmodulin-dependent protein kinase II (CaMKII), calmodulin (CaM) mRNA, and cAMP-response element binding protein (CREB) mRNA in the hippocampus of rats. METHODS: The rats were divided randomly into stressed and control groups. The stressed group was given chronic multiple-stress for 6 weeks to set up a chronic multiple-stressed model. The rats' performance of spatial learning and memory was tested using Morris Water Maze (MWM) and Y-maze. Meanwhile, the expressions of CaMKII, CaM mRNA and CREB mRNA of rats' hippocampus were detected by immunohistochemistry, Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. In addition, the width of synaptic cleft and the thickness of post-synaptic densities (PSD) were observed in the hippocampal CA3 region of rats by electron microscopy. RESULTS: After exposure to chronic multiple-stress for 6 weeks, the ability of learning and memory of the stressed group was higher than that of the control group (P < 0.05, P < 0.01). The width of synaptic cleft was smaller and the thickness of PSD was larger in the hippocampal CA3 region of the stressed group than in that of the control group (P < 0.01). The CaMK II immunostaining of the stressed group was stronger than that of the control group in the stratum radiatum and oriens of the hippocampal CA1 and CA3, especially in the stratum oriens. Quantitative analysis indicated that the expression of CaMK II, CaM mRNA, and CREB mRNA in the hippocampus of the stressed group was higher than that of the control group (P < 0.05, P < 0.01). CONCLUSIONS: The capacity of learning and memory can be enhanced after chronic multiple-stress. The increased levels of CaMK II, CaM mRNA, and CREB mRNA may contribute to the enhancing effect of chronic multiple-stress on learning and memory.

[Study on the characterization of the bi-directional promoter between pp38 gene and 1.8kb mRNA transcripts of Marek's disease viruses].

The bi-directional promoter between pp38 gene and 1.8kb mRNA transcripts of Marek's disease viruses (MDV) was divided into two single-direction promoters from the replication of MDV genomic DNA. The pp38 gene was cloned into pUC18 vector for plasmid pUC-pp38. Then the complete bi-directional promoter was cloned into pUC-pp38 in two directions to form plasmids pPro(f)pp38 and pPro(r)pp38, and the divided two single directional promoters were cloned in pUC-pp38 for plasmids pdPro(f)pp38 and pdPro(r)pp38. 24 to 48 hours after transfection to chicken embryo fibroblast (CEF) cells, the expression of pp38 could be detected in above 4 samples with Indirect Immuno-fluorescent Assay (IFA). In order to analysis the activity of the promoter quantificationally, CAT was used as the report gene. The complete or divided promoters were cloned into pCAT-Basic vector for plasmids pPro(f)CAT, pPro(r)CAT, pdPro(f)CAT and pdPro(r)CAT. The activity of CAT was measured from the lysed CEF cells, when they were transfected for 48 hours by the above four plasmids, respectively. The results showed the activity of the divided promoters reduce on both directions, especially for the direction of 1.8kb mRNA transcript, nearly down to 1/41.

[Risk factors for stroke in Chinese with non valvular atrial fibrillation: a case-control study].

OBJECTIVE: The aim of this study is to identify the risk factors in Chinese with nonvalvular atrial fibrillation and stroke, using case-control methodology. METHODS: A total of 4 511 adult patients diagnosed with atrial fibrillation were identified from 18 hospitals over a 2-year period. There were 1 086 patients with rheumatic valvular atrial fibrillation and 3 425 patients with nonvalvular atrial fibrillation. Among the nonvalvular atrial fibrillation patients, 827 had ischemic stroke. The data of the patients having nonvalvular atrial fibrillation with stroke was compared with those having nonvalvular atrial fibrillation without stroke (n = 2 598). The effect of each variable on stroke was assessed with a logistic regression analysis. RESULTS: The studied cases with stroke and controls without stroke were similar in terms of percentage with sex, a past history of congestive heart failure, myocardial infarction, and mean left atrial size. Cases were significantly older than controls (73.3 +/- 9.2 vs. 68.2 +/- 12.3, P < 0.001) and more likely to have a history of hypertension (71.0% versus 51.6%, P < 0.001) and diabetes (17.9% vs. 11.1%, P = 0.001). There is strong evidence that left atrial (LA) thrombi make AF patients highly risky for stroke. In multivariate analysis, age > or = 75 (OR 1.76; 95% CI 1.08 approximately 2.98), history of hypertension (OR 1.52; 95% CI 1.28 approximately 1.80), diabetes (OR 1.39; 95% CI 1.11 approximately 1.76), high systolic blood pressure (OR 1.71; 95% CI 1.21 approximately 2.28), LA thrombi (OR 2.77; 95% CI 1.25 approximately 6.13) were independently associated with stroke. The lack of the association between left ventricular dysfunction and stroke is due to the relatively incorrect diagnosis of heart failure in the context of atrial fibrillation. CONCLUSIONS: Our analysis suggests that old age, hypertension, diabetes, high systolic blood pressure and LA thrombi detected with echocardiography are independent risk factors, which should be considered when decision of long-term anticoagulation therapy to prevent stroke with nonvalvular atrial fibrillation is to be made.