Transcriptional analysis of porcine circovirus-like virus P1.

BACKGROUND: Recently identified porcine circovirus-like virus P1 has the smallest DNA viral genome. In this study, we identified the viral genes and their corresponding mRNA transcripts. RESULTS: The RNAs of P1, synthesized in porcine kidney cells, were examined with northern blotting and PCR analyses. Eight virus-specific RNAs were detected. Four mRNAs (open reading frames (ORFs) 1, 2, 4, and 5) are encoded by the viral (-) strand and four (ORFs 3, 6, 7, and 8) are encoded by the viral (+) strand. All proteins encoded by the ORFs of the P1 virus are less than 50 amino acids in length, except that encoded by ORF1 (113 amino acids). CONCLUSIONS: We show a very complex viral transcription pattern in P1-infected cells.

[Epidemiological study on developmental delay of 18-month-old children from four districts/counties in Beijing].

OBJECTIVE: To investigate epidemiological characteristics of prevalence, impact factors and etiology on developmental delay of 18-month-old children from four districts/counties in Beijing. METHODS: An epidemiological study on developmental delay was designed to investigate all the 18-month-old children enrolled from Shunyi,Daxing,Miyun and Yanqing districts/counties in Beijing from May to September, 2011. Combining the tertiary network of child health with hospital clinical study was used. Child developmental questionnaires were completed by doctors in communities of the first network of child health. Gesell Developmental Schedules for children with Denver developmental screening test (DDST) screening positive results were assessed by doctors in districts/counties hospitals of the second network of child health. The children diagnosed as developmental delay were transferred to the tertiary hospitals of the third network of child health for further etiological diagnosis, follow-up and developmental evaluation. The case-control study compared between children with/without developmental delay were performed in accordance with the 1:4 ratios by gender and residence community matched. SPSS 16.0 was adopted for data analysis of the case-control study. RESULTS: A total of 3 182 children were screened among the 4 037 children fitting the criteria,and the coverage rate was 78.8% (3 182/4 037). Of the 3 182 screened children, 22 children were diagnosed as developmental delay. The prevalence rate was 6.91 ‰ (22/3 182). Out of the 22 children with developmental delay, 15 were boys and 7 were girls. The sex ratio was 2.1:1. The prevalence rates of the children with developmental delay in Shunyi, Daxing, Miyun and Yanqing were 3.45 ‰ (4/1 160), 4.50 ‰(5/1 111), 15.87 ‰(7/441) and 12.77 ‰ (6/479), respectively. The results from one-way ANOVA analysis showed the main risk factors in children with developmental delay included low-income families, mothers' low educational level, small size for gestational age infant, multiple fetuses, serious diseases after birth, congenital malformations and physical retardation (P<0.05). CONCLUSION: The screening coverage rate of this study is 78.8%. The prevalence rate of children with developmental delay is 6.91 ‰, which is significantly different in sex ratio and districts of the subjects. The etiology of developmental delay might be associated with social-economic and biological factors.

Genome-Wide Analysis of the Soybean TIFY Family and Identification of GmTIFY10e and GmTIFY10g Response to Salt Stress.

TIFY proteins play crucial roles in plant abiotic and biotic stress responses. Our transcriptome data revealed several TIFY family genes with significantly upregulated expression under drought, salt, and ABA treatments. However, the functions of the GmTIFY family genes are still unknown in abiotic stresses. We identified 38 GmTIFY genes and found that TIFY10 homologous genes have the most duplication events, higher selection pressure, and more obvious response to abiotic stresses compared with other homologous genes. Expression pattern analysis showed that GmTIFY10e and GmTIFY10g genes were significantly induced by salt stress. Under salt stress, GmTIFY10e and GmTIFY10g transgenic Arabidopsis plants showed higher root lengths and fresh weights and had significantly better growth than the wild type (WT). In addition, overexpression of GmTIFY10e and GmTIFY10g genes in soybean improved salt tolerance by increasing the PRO, POD, and CAT contents and decreasing the MDA content; on the contrary, RNA interference plants showed sensitivity to salt stress. Overexpression of GmTIFY10e and GmTIFY10g in Arabidopsis and soybean could improve the salt tolerance of plants, while the RNAi of GmTIFY10e and GmTIFY10g significantly increased sensitivity to salt stress in soybean. Further analysis demonstrated that GmTIFY10e and GmTIFY10g genes changed the expression levels of genes related to the ABA signal pathway, including GmSnRK2, GmPP2C, GmMYC2, GmCAT1, and GmPOD. This study provides a basis for comprehensive analysis of the role of soybean TIFY genes in stress response in the future.

Genome-Wide Analysis of DEAD-box RNA Helicase Family in Wheat (Triticum aestivum) and Functional Identification of TaDEAD-box57 in Abiotic Stress Responses.

DEAD-box RNA helicases constitute the largest subfamily of RNA helicase superfamily 2 (SF2), and play crucial roles in plant growth, development, and abiotic stress responses. Wheat is one of the most important cereal crops in worldwide, and abiotic stresses greatly restrict its production. So far, the DEAD-box RNA helicase family has yet to be characterized in wheat. Here, we performed a comprehensive genome-wide analysis of the DEAD-box RNA helicase family in wheat, including phylogenetic relationships, chromosomal distribution, duplication events, and protein motifs. A total of 141 TaDEAD-box genes were identified and found to be unevenly distributed across all 21 chromosomes. Whole genome/segmental duplication was identified as the likely main driving factor for expansion of the TaDEAD-box family. Expression patterns of the 141 TaDEAD-box genes were compared across different tissues and under abiotic stresses to identify genes to be important in growth or stress responses. TaDEAD-box57-3B was significantly up-regulated under multiple abiotic stresses, and was therefore selected for further analysis. TaDEAD-box57-3B was localized to the cytoplasm and plasma membrane. Ectopic expression of TaDEAD-box57-3B in Arabidopsis improved tolerance to drought and salt stress as measured by germination rates, root lengths, fresh weights, and survival rates. Transgenic lines also showed higher levels of proline and chlorophyll and lower levels of malonaldehyde (MDA) than WT plants in response to drought or salt stress. In response to cold stress, the transgenic lines showed significantly better growth and higher survival rates than WT plants. These results indicate that TaDEAD-box57-3B may increase tolerance to drought, salt, and cold stress in transgenic plants through regulating the degree of membrane lipid peroxidation. This study provides new insights for understanding evolution and function in the TaDEAD-box gene family.

[Myocardial damage and change of mitochondrial Mn-superoxide dismutase activity in craniocerebral injured rats and the intervening effect of ginkgo biloba extract].

OBJECTIVE: To investigate the myocardial damage and changes of myocardial mitochondrial Mn-superoxide dismutase (Mn-SOD) activity in craniocerebral injured rats and the effect of Ginkgo biloba extract (GBE) on them. METHODS: Craniocerebral injured rats model was established by fluid-percussion and treated with GBE. The dynamical changes of electrocardiograph (ECG) in 24 h were monitored, the serum level of MB isoenzyme of creatine kinase (CK-MB) and the change of myocardial mitochondrial Mn-SOD activity as well as the pathologic changes of myocardium (HE staining) were observed. RESULTS: The occurrence of ECG abnormality obviously increased in the injured rats, accompanied with increased serum CK-MB (P<0.05) and decreased myocardial Mn-SOD levels (P<0.05), and the Mn-SOD activity was negatively correlated with the level of CK-MB (r=-0.997, P<0.05). Pretreatment of GBE resulted in the decrease of ECG abnormality occurrence (P<0.01), serum CK-MB level (P<0.05), and degree of myocardial damage, as well as the increase of Mn-SOD activity in post-craniocerebral injured rats. CONCLUSIONS: Craniocerebral injury can result in distinct myocardial damage, which is possibly correlated with the lowering of anti-oxidation stress level of myocardial cellular mitochondria. GBE possesses the protective effect on myocardial damage after craniocerebral injury.

Effect of cognitive behavioral therapy for heart failure: A systematic review protocol of randomized controlled trial.

BACKGROUND: This proposed study will systematically assess the effect and safety of cognitive-behavioral therapy (CBT) for heart failure (HF). METHODS: We will search the following electronic databases for randomized controlled trials assessing the effect of CBT in patients with HF: PUBMED, EMBASE, Cochrane Library, Web of Science, Scopus, Chinese Biomedical Literature Database, China National Knowledge Infrastructure, VIP Information, and Wanfang Data from their inceptions to present without any language limitations. Two authors will independently conduct the study selection, data extraction, and methodological quality assessment. The methodological quality will be evaluated by Cochrane risk of bias tool. RESULTS: This study will assess the efficacy and safety of CBT for patients with HF. The primary outcomes consist of depression and anxiety. The secondary outcomes comprise of all-cause mortality, change in body weight, urine output, change in serum sodium; and any adverse events. CONCLUSION: The results of this study will summarize the up-to-date evidence on the effect and safety of CBT for HF. PROSPERO REGISTRATION NUMBER: PROSPERO CRD42019135932.

[Comparative study with the effect of stromal cell derived factor-1 on osteogenic differentiation of human healthy and inflammatory periodontal ligament stem cells].

OBJECTIVE: This study aims to compare the osteogenic differentiation capability of stem cells derived from human inflammatory periodontal ligament tissues (iPDLSCs) with those of stem cells derived from healthy periodontal ligament tissues (hPDLSCs). Both types of tissues were induced by stromal cell derived factor (SDF-1) in vitro. METHODS: iPDLSCs and hPDLSCs were primarily cultured by tissue digestion method and purified by limited dilution cloning. The cells were passaged and identified by stem cell surface marker expression through flow cytometry. Then, we used thiazolyl blue tetrazolium bromide to detect and compare the proliferation capabilities of the iPDLSCs and hPDLSCs. Express of bone volumes were detected by alizarin red staining after SDF-1 was added to the cells. Using alkaline phosphatase, we evaluated the osteogenic differentiation capability of the cells induced by SDF-1. The expression levels of the osteogenesis-related genes of the cells induced by SDF-1 were determined by reverse transcription-polymerase chain reaction. RESULTS: After purification, both iPDLSCs and hPDLSCs expressed stem cell markers. hPDLCSs had a higher proliferation capability than iPDLSCs. Osteogenesis-related genes had higher expression levels in the cells induced by SDF-1 than in those without induction (P<0.05). SDF-1 at 50 and 200 ng·mL⁻¹ concentration greatly affected the differen-tiation capabilities of iPDLSCs and hPDLSCs respectively. CONCLUSIONS: iPDLSCs and hPDLSCs had osteogenic differentia-tion capability. The level of osteogenic differentiation in normal and inflamed periodontal ligament stem cells increases after SDF-1 induction.

[Recombinant human growth hormone promotes in vitro mesenchymal stem cell growth].

OBJECTIVE: To detect the effect of recombinant human growth hormone (rhGH) to mesenchymal stem cells (MSCs) in vitro. METHODS: Various concentrations of rhGH (terminal concentrations 1, 10, 50, 100, 200 and 500 microg/L) were added to medium containing the second generation of MSCs, A value was measured by MTT method at various concentrations and at 24, 48 and 72 h. The expression of insulin-like growth factor 1(IGF-1) mRNA in the presence (24, 48 and 72 h) or absence (the 4(th) day, the 9(th) day, the 10(th) day, the 2(nd) week, the 3(rd) week and the 4(th) week post removal of rhGH) of various rhGH concentrations was determined by RT-PCR. RESULTS: rhGH could promote the MSCs growth at concentration > 10 microg/L in a time-dependent manner. The optimal concentration was 200 microg/L with a growth rate 144.74%. The expression of IGF-1 mRNA increased in a time-dependent manner gradually (0.6749 +/- 0.0084, 0.7781 +/- 0.0068, 0.8230 +/- 0.0060 at 24, 48 and 72 h, respectively at 200 microg/L rhGH). After rhGH withdraw, the expression of IGF-1 mRNA decreased in a time-dependent manner till 2 weeks (0.5287 +/- 0.0077, 0.5747 +/- 0.0050, 0.6068 +/- 0.0056, 0.7071 +/- 0.0089, 0.5791 +/- 0.0057, 0.5781 +/- 0.0081 at the 4(th) day, the 9(th) day, the 10(th) day, the 2(nd) week, the 3(rd) week and the 4(th) week post removal of rhGH). CONCLUSION: rhGH could promote the growth of MSCs in vitro.

[Therapeutic effects of recombinant human growth hormone and mesenchymal stem cells transplantation alone or in combination in adriamycin induced cardiomyopathy rat model].

OBJECTIVE: To investigate the therapeutic effects of recombinant human growth hormone (rhGH) and mesenchymal stem cells (MSCs) transplantation alone or in combination in adriamycin-induced cardiomyopathy (CM) rat model. METHODS: Wistar rats were divided into normal (n = 6), CM (n = 9), rhGH (2 mg/kg, subcutaneous injection, qod for 1 month, n = 6), MSCs transplantation (MSCs group, 5 x 10(6)/100 microl, n = 7) and rhGH + MSCs (G + M group, n = 7) groups. After 1 month, hemodynamic data, the ratio of heart weight/body weight (HW/BW) and left ventricular weight/body weight (LW/BW) were obtained. The myocardial MSCs marked by BrdU were detected by immunohistochemistry. Cardiac myosin heavy-chain (MHC) and its isozymes were detected by SDS-PAGE technique. Serum growth hormone (GH) and B-type natriuretic peptide (BNP) levels were measured. RESULTS: Compared to CM group, cardiac function remained unchanged in MSC group while significantly improved in rhGH group and further improved in G + M group. HW/BW and LW/BW were similar among groups (all P > 0.05). The MSCs number in the myocardium of G + M group was significantly higher than those of MSCs group. Compared to CM group, V3-MHC was significantly increased in MSCs group (10.33 +/- 0.33 vs 7.43 +/- 2.08, P < 0.05), V1 and V3-MHC were significantly increased in G + M group (86.22 +/- 1.73 vs 77.47 +/- 2.02, 12.44 +/- 0.31 vs 7.43 +/- 2.08, all P < 0.05). GH levels were significantly higher in rhGH and G + M treated animals than that in CM animals [(2.50 +/- 0.68) vs (1.37 +/- 0.09) microg/L, (2.48 +/- 0.90) vs (1.25 +/- 0.42) microg/L, all P < 0.05], BNP levels were significantly lower in rhGH, MSCs and G + M treated groups than that in CM group [(1270.72 +/- 203.72) vs (1462.44 +/- 242.87) ng/L, (1385.00 +/- 250.13) vs ( 1475.29 +/- 281.33) ng/L, (1219.31 +/- 126.71) vs (1451.78 +/- 180.93) ng/L, all P < 0.05]. CONCLUSIONS: rhGH but not MSC could improve the cardiac function in adriamycin-induced CM rats. Combined rhGH and MSCs transplantation therapy could further improve cardiac function possibly due to enhanced MSCs growth and transformation into cardiomyocytes.

Physicochemical properties of corn stalk after treatment using steam explosion coupled with acid or alkali.

The aim of this study was to evaluate comparatively the effects of different pretreatments including steam explosion, acid, and alkali, alone or in combination, on the structural properties and thermal stability of corn stalk. All of the treated treatments decreased the contents of hemicellulose and lignin and thereby increased the content of cellulose in corn stalks. But the combined treatments with alkali and steam explosion under 0.4-0.6 MPa were better as compared with other treatments based on the removals of hemicellulose and lignin, and about 71.58-79.59% of hemicellulose and 64.32-71.83% of lignin were removed. Treatment with steam explosion coupled with acid or alkali changed the bonding distribution and surface morphology and increased the crystallinity and thermal stability of corn stalks, and the degradation temperature reached over 350 °C. These results suggest that steam explosion coupled with alkali is a better method for the depolymerization of corn stalk polymer.

Driver sleepiness and risk of car crashes in Shenyang, a Chinese northeastern city: population-based case-control study.

OBJECTIVE: To estimate the association of driver sleepiness with the risk of car crashes. METHODS: A population-based case-control study was conducted in Shenyang, a northeastern city in China, between November 2001 and July 2002. The case group comprised 406 car drivers involved in crashes, and 438 car drivers recruited at randomly selected sites, and on the day of week, and the time of day when they were driving on highways in the study region during the study period were used as control groups. Face-to-face interviews with drivers were conducted according to a well-structured questionnaire covering the circumstances of their current trip and their background information. Stanford sleepiness scale and Epworth sleepiness scale were used to quantify acute sleepiness and chronic sleepiness respectively. RESULTS: There was a strong association between chronic sleepiness and the risk of car crash. Significantly increased risk of crash was associated with drivers who identified themselves as sleepy (Epworth sleepiness score > or = 10 vs < 10; adjusted odds ratio 2.07, 95% confidence interval 1.30 to 3.29), but no increased risk was associated with measures of acute sleepiness. CONCLUSIONS: Chronic sleepiness in car drivers significantly increases the risk of car crash. Reductions in road traffic injuries may be achieved if fewer people drive when they are sleepy.

Recombinant human growth hormone secreted from tissue-engineered bioartificial muscle improves left ventricular function in rat with acute myocardial infarction.

BACKGROUND: Experimental studies and preliminary clinical studies have suggested that growth hormone (GH) treatment may improve cardiovascular parameters in chronic heart failure (CHF). Recombinant human GH (rhGH) has been delivered by a recombinant protein, by plasmid DNA, and by genetically engineered cells with different pharmacokinetic and physiological properties. The present study aimed to examine a new method for delivery of rhGH using genetically modified bioartificial muscles (BAMs), and investigate whether the rhGH delivered by this technique improves left ventricular (LV) function in rats with CHF. METHODS: Primary skeletal myoblasts were isolated from several Sprague-Dawley (SD) rats, cultured, purified, and retrovirally transduced to synthesize and secrete human rhGH, and tissue-engineered into implantable BAMs. Ligation of the left coronary artery or sham operation was performed. The rats that underwent ligation were randomly assigned to 2 groups: CHF control group (n = 6) and CHF treatment group (n = 6). The CHF control group received non-rhGH-secreting BAM (GFP-BAMs) transplantation, and the CHF treatment group received rhGH-secreting BAM (GH-BAMs) transplantation. Another group of rats served as the sham operation group, which was also randomly assigned to 2 subgroups: sham control group (n = 6) and sham treatment group (n = 6). The sham control group underwent GFP-BAM transplantation, and the sham treatment group underwent GH-BAM transplantation. GH-BAMs and GFP-BAMs were implanted subcutaneously into syngeneic rats with ligation of the left coronary artery or sham operation was performed. Eight weeks after the treatment, echocardiography was performed. hGH, insulin-like growth factor-1 (IGF-1) and TNF-alpha levels in rat serum were measured by radioimmunoassay and ELISA, and then the rats were killed and ventricular samples were subjected to immunohistochemistry. RESULTS: Primary rat myoblasts were retrovirally transduced to secrete rhGH and tissue-engineered into implantable BAMs containing parallel arrays of postmitotic myofibers. In vitro, they secreted 1 to 2 microg of bioactive rhGH per day. When implanted into syngeneic rat, GH-BAMs secreted and delivered rhGH. Eight weeks after therapy, LV ejection fraction (EF) and fractional shortening (FS) were significantly higher in CHF rats treated with GH-BAMs than in those treated with GFP-BAMs ((65.0 +/- 6.5)% vs (48.1 +/- 6.8)%, P < 0.05), ((41.3 +/- 7.4)% vs (26.5 +/- 7.1)%, P < 0.05). LV end-diastolic dimension (LVEDD) was significantly lower in CHF rats treated with GH-BAM than in CHF rats treated with GFP-BAM (P < 0.05). The levels of serum GH and IGF-1 were increased significantly in both CHF and sham rats treated with GH-BAM. The level of serum TNF-alpha decreased more significantly in the CHF treatment group than in the CHF control group. CONCLUSIONS: rhGH significantly improves LV function and prevents cardiac remodeling in rats with CHF. Genetically modified tissue-engineered bioartificial muscle provides a method delivering recombinant protein for the treatment of heart failure.

[Blood lead level and related risk factors among children aged 0-6 years in Beijing].

OBJECTIVE: To observe blood lead level and related risk factors among children of 0 - 6-year old in Beijing. METHODS: Stratified-clustered-random sampling and simple random sampling were used. A total of 2 262 children of 0 - 6 years old were investigated from May to July 2001. They were permanent residents in Beijing. Blood lead level was tested by graphite atomizer absorption spectrophotometer. At the same time, related factors were investigated using a standardized questionnaire. RESULTS: The mean lead level of children in Beijing was 96.8 micro g/L with 35.7% of those >/= 100 micro g/L, and 2.5% >/= 200 micro g/L. The proportion of lead poisoning was significantly higher in rural children (P < 0.01). The proportion of lead poisoning in 2-year-old group was higher than that in other age groups. The proportion of lead poisoning in boys was significantly higher than that in girls (P < 0.01). The following factors might serve as major risk factors related to child lead poisoning, such as, never or rarely drinking milk, living in nearby highways (less than 50 meters) or living in the first floor/bungalow, and so on. CONCLUSION: The blood lead level of >/= 100 micro g/L among Beijing children appeared to be a big problem. Decision-makers should pay more attention to prevent blood lead level being high, and to cure these children who suffered in lead poisoning. Effective intervention measures on these target populations should be taken.

[A case-control study on the risk factors for road injury].

OBJECTIVE: To investigate the risk factors of road injury. METHODS: Case-control study was used. From November 2001 to August 2002, 406 drivers who had 438 drivers who had not experienced a motor vehicle crash in Huanggu district, Shenyang city were recruited by randomly selection on time of day, day of week and site in the same period at same district. Face to face interviews with drivers were conducted according to a highly structured questionnaire covering the circumstances of the current trip, usual behavior and background characteristics of the drivers and the condition of motor vehicles. Stanford sleepiness scale and Epworth sleepiness scale were used to quantify acute and chronic sleepiness respectively. RESULTS: Increased risk was associated with drivers who identified themselves as having chronic doziness (OR = 1.98, 95% CI: 1.26 - 3.12). Increase in risk was associated with measures of acute tiredness, but without statistical significance (OR = 2.38, 95% CI: 0.89 - 6.31). Comparing to permanent daytime work pattern, rotating shifts or permanent night-work pattern increased the risk of crash (OR = 2.09, 95% CI: 1.48 - 2.94). The risk of motor vehicle crash among the drivers who drank alcohol in the previous 6 hours was 3.59 times (95% CI: 1.13 - 11.39) of those drivers who did not drink. Driving violations also contributed to the increased risk of crash (OR = 1.73, 95% CI: 1.22 - 2.46). CONCLUSION: Factors as chronic doziness, rotating shifts or permanent night-work pattern, driving under alcohol impairment, violation of motor vehicle regulation all significantly increased the risk of road injury. Acute sleepiness might serve as a potential risk factor for road injury.