Puerarin alleviates the progression of non-small cell lung cancer by regulating the miR-342/CCND1 axis.

Puerarin has recently been demonstrated to play anti-cancer roles in a series of human cancers, including non-small cell lung cancer (NSCLC), possibly through regulation of cancer-related microRNAs (miRNAs). The purpose of the present study was to further investigate the detailed role and underlying mechanism of puerarin on NSCLC progression. Cell viability and apoptosis were assessed using the Cell Counting kit-8 (CCK-8) assay and flow cytometry, respectively. Transwell assays were performed to determine cell migration and invasion abilities. The qRT-PCR assay was employed to detect the expression of miR-342 and cyclin D1 (CCND1) mRNA, and CCND1 protein expression was evaluated by western blotting. The targeted interaction between miR-342 and CCND1 was verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. We found that our data demonstrated that puerarin repressed cell viability, migration, invasion, and cell cycle progression, and enhanced the apoptosis of NSCLC cells. miR-342 overexpression hindered the migration, invasion and cell cycle progression, and accelerated the apoptosis of NSCLC cells. miR-342 inhibited CCND1 expression by directly binding to the 3'-UTR of CCND1. Moreover, miR-342 overexpression-mediated anti-migration, anti-invasion, anti-cell cycle progression, and pro-apoptotic effects were abated by co-transfection of pcDNA-CCND1. More importantly, puerarin inhibited CCND1 expression by upregulating miR-342. Additionally, puerarin hampered NSCLC cell progression in vitro and tumor growth in vivo by upregulating miR-342. In conclusion, our study suggested that puerarin hampered NSCLC progression in vitro and in vivo at least partly through regulating miR-342/CCND1 axis, highlighting a novel mechanism of puerarin exerting anti-cancer property in NSCLC.

Effects of dietary energy levels on physiological parameters and reproductive performance of gestating sows over three consecutive parities.

OBJECTIVE: This experiment was to evaluate the effects of the dietary energy levels on the physiological parameters and reproductive performance during gestation over three parities in sows. METHODS: A total of 52 F1 gilts (Yorkshire×Landrace) were allotted to one of four dietary treatments using a completely randomized design. The treatments contained 3,100, 3,200, 3,300, or 3,400 kcal of metabolizable energy (ME)/kg diet but feed was provided at 2.0, 2.2, and 2.4 kg/d in the first, second and third parity, respectively. RESULTS: The body weight and body weight gain during gestation increased as the dietary energy level increased (p<0.05, and p<0.01) in the first parity. In the second parity, the body weight of sows was the lowest (p<0.05) when 3,100 kcal of ME/kg treatment diet was provided. The body weight was higher as the dietary energy level increased (p<0.05) during the gestation period in the third parity. During lactation, the voluntary feed intake of lactating sows tended to decrease when gilts were fed higher energy treatment diet (p = 0.08) and the body weight, body weight gain were increased by dietary energy level during gestation (p< 0.05). Backfat thickness was not affected by dietary treatment during the gestation period in three parities, interestingly backfat change from breeding to d 110 of gestation was higher as the dietary energy level increased at the first parity (p<0.05). When gilts were fed 3,400 kcal of ME/kg treatment diet a higher number of weaning piglets was observed in the first parity (p<0.05). The highest culling rate (69%) was seen when gestating sows were fed 3,100 kcal/kg ME treatment diet during three parities. CONCLUSION: In conclusion, the adequate energy intake of gestating sows should be 6,400 or 6,600 kcal of ME/d, 7,040 or 7,260 kcal of ME/d, and 7,680 or 7,920 kcal of ME/d for parity 1, 2, and 3, respectively.

Effect of rapeseed meal supplementation to gestation diet on reproductive performance, blood profiles and milk composition of sows.

OBJECTIVE: This experiment evaluated the effect of dietary supplementation levels of rapeseed meal (RSM) in gestation diets on reproductive performance, blood profiles, milk composition of sows, and growth of their progeny. METHODS: A total of 55 mixed-parity sows (Yorkshire×Landrace; average parity = 3.82) with an initial body weight (BW) of 193.0 kg were used in this experiment. Sows were allotted to one of 5 treatments at breeding based on BW and backfat thickness in a completely randomized design. Treatments consisted of dietary RSM supplementation levels (0%, 3%, 6%, 9%, and 12%) in gestation diets. During lactation all sows were fed a common lactation diet with no RSM supplementation. RESULTS: Body weight, backfat thickness, litter size, lactation feed intake, and milk composition of sows, and growth of their progeny were not different among dietary treatments. In blood profiles, a quadratic increase (Quadratic, p<0.05) in serum triiodothyronine (T3) concentration and a linear increase (Linear, p<0.01) in serum thyroxine (T4) concentration were observed at d 110 of gestation as dietary RSM supplementation levels increased. However, serum T3 and T4 concentrations in lactating sows and their piglets were not affected by RSM supplementation of gestation diets. Concentrations of serum total cholesterol and low density lipoprotein cholesterol in sows were not influenced by dietary treatments, whereas serum glucose level in sows decreased linearly at d 110 of gestation (Linear, p<0.05) by increasing dietary RSM supplementation in gestation diets. CONCLUSION: The RSM could be supplemented to gestation diets up to 12% with no detrimental effects on reproductive performance and growth of their progeny. However, increasing supplementation levels of RSM in gestation diets may increase serum T3 and T4 concentrations and decrease serum glucose concentration of sows in late gestation.

[Analysis on the novel compound heterozygous mutation Fâ ª of a patient with hereditary factor â ª deficiency].

Objective: To investigate the clinical phenotype and genotype characteristics of a Chinese hereditary factor Ⅺ deficiency pedigree. Methods: The activated partial thromboplastin time (APTT), prothrombin time (PT), FⅪ activity (FⅪ: C) were measured by clotting method using automatic coagulation analyzer. The FⅪ antigen (FⅪ: Ag) was assayed by enzyme-linked immunosorbent assay (ELISA). Fifteen exons of F11 from the proband and his pedigree members were amplified by polymerase chain reaction (PCR), then sequenced. Pymol software was used to analyze the novel mutations. Results: APTT, FⅪ: C and FⅪ: Ag of proband was 74.2 s, 4.0% and 2.9%, respectively. For his older sister, APTT, FⅪ: C and FⅪ: Ag was 67.1 s, 3.0% and 1.8%, respectively. APTT, FⅪ: C and FⅪ: Ag of healthy controls were 34.5 s, 100.0% and 100.0%. FⅪ: C of proband's father, mother and brother were 72.0%, 62.0%, and 78.0%, respectively. FⅪ: Ag of them were 50.0%, 43.0%, and 51.8%, respectively. The other coagulant parameters of the proband and his pedigree were all in the normal range. Sequence analysis showed two heterozygous gene mutations in F11 of the proband and his older sister. One was a deletion of T at nucleotide 1 491 in exon 12, resulting in a frameshift. A substitution of leucine 465 by tryptophan and a terminal coden after 7 amino acid: F11NM_13142c.1491delT (p.Leu465Trp.fs*7). The other was a G to A substitution at nucleotide 1 815 in exon 15, resulting in a substitution of glycine 573 by aspartic acid: F11 NM_13142c.1815G>A (p.Gly573Asp). F11NM_13142c.1491delT (p.Leu465Trp.fs*7) heterozygotes were found both in the proband's father and his brother while p. Gly573Asp heterozygote was only found in his mother. F11 of the proband's uncle was wild. Conclusion: The novel compound heterozygous mutations of F11NM_13142c.1491delT (p.Leu465Trp.fs*7) and F11 NM_13142 c. 1815G>A (p.Gly573Asp) are responsible for FⅪ deficiency to the proband, which induced the decrease of FⅪ: C and FⅪ: Ag.

Effects of Dietary Energy Levels on the Physiological Parameters and Reproductive Performance of Gestating Gilts.

This experiment was conducted to investigate the effects of dietary energy levels on the physiological parameters and reproductive performance of gestating first parity sows. A total of 52 F1 gilts (Yorkshire×Landrace) were allocated to 4 dietary treatments using a completely randomized design. Each treatment contained diets with 3,100, 3,200, 3,300, or 3,400 kcal of metabolizable energy (ME)/kg, and the daily energy intake of the gestating gilts in each treatment were 6,200, 6,400, 6,600, and 6,800 kcal of ME, respectively. During gestation, the body weight (p = 0.04) and weight gain (p = 0.01) of gilts linearly increased with increasing dietary energy levels. Backfat thickness was not affected at d110 of gestation by dietary treatments, but increased linearly (p = 0.05) from breeding to d 110 of gestation. There were no significant differences on the litter size or litter birth weight. During lactation, the voluntary feed intake of sows tended to decrease when the dietary energy levels increased (p = 0.08). No difference was observed in backfat thickness of the sows within treatments; increasing energy levels linearly decreased the body weight of sows (p<0.05) at d 21 of lactation and body weight gain during lactation (p<0.01). No significant differences were observed in the chemical compositions of colostrum and milk. Therefore, these results indicated that high-energy diets influenced the bodyweight and backfat thickness of sows during gestation and lactation. NRC (2012) suggested that the energy requirement of the gestation gilt should be between 6,678 and 7,932 kcal of ME/d. Similarly, our results suggested that 3,100 kcal of ME/kg is not enough to maintain the reproductive performance for gilts during gestation with 2 kg feed daily. Gilts in the treatment 3,400 kcal of ME/kg have a higher weaning number of piglets, but bodyweight and backfat loss were higher than other treatments during lactation. But bodyweight and backfat loss were higher than other treatments during lactation. Consequently, an adequate energy requirement of gestating gilts is 6,400 kcal of ME/d.

The Effects of Gilts Housed Either in Group with the Electronic Sow Feeding System or Conventional Stall.

This experiment was conducted to assess the welfare and productivity of gestating gilts in groups with the electronic sow feeding (ESF) system compared to conventional stalls. A total of 83 gilts (Yorkshire×Landrace) were housed into individual stalls to be artificially inseminated. Gilts confirmed pregnant were introduced to their treatment, conventional stalls (ST) or groups with the ESF system. All gilts were taken to the farrowing crates one week prior to their expected farrowing date. In the gestation period, there were no significant differences between gilts allocated to ST and ESF on growth performance. However, backfat thickness gain (p = 0.08) and body condition score (BCS) at 110 days of gestation (p = 0.10) tended to be higher in ESF gilts than ST. Likewise, gilts housed in group showed significantly higher estimated body muscle contents at 110 days of gestation (p = 0.02) and body muscle change during gestation (p = 0.01). There was a trend for a shorter parturition time in ESF gilts (p = 0.07). In the lactation period, group housed gilts showed a tendency to increased BCS changes (p = 0.06). Reproductive performance did not differ with the exception of piglet mortality (ST = 0.2 no. of piglets vs ESF = 0.4 no. of piglets; p = 0.01). In blood profiles, ST gilts showed a higher cortisol level at 110 days of gestation (p = 0.01). Weaning to estrus interval was shorter in gilts housed in ESF than ST (p = 0.01). In locomotory behaviors, ESF gilts recorded a tendency to elevate locomotion score at 36, 70, and 110 days of gestation (p = 0.07, p = 0.06, and p = 0.06, respectively). Similarly, ESF gilts showed significantly higher incidence of scratches at 36, 70, and 110 days of gestation (p = 0.01). Moreover, farrowing rates were higher in stall treatment (97.6%) compare to group housing treatment (95.2%). In conclusion, while group housed gilts with ESF system positively affected welfare status in combination with less physiologically stressful environments and activity, it negatively effects piglet mortality, farrowing rates and injuries of gilts.

[A cross-sectional study on AIDS-related knowledge and demands for health education among first-grade students of senior high schools in Tianjin].

Objective: To understand the AIDS awareness and related factors as well as the demands for AIDS-related health education programs, among first-grade students from three senior high schools in Tianjin. Methods: A cross-sectional study using both stratified cluster and convenient sampling methods was conducted. Anonymous questionnaires including social demographic characteristics, AIDS-related knowledge and needs for AIDS health education were distributed to the first-grade students from two general middle schools and one vocational senior middle school in Tianjin. Results: A total of 1 082 students were responding to the questionnaires. Among them, the overall awareness rates of AIDS-related knowledge were 34.3%(371/1 082), 71.9%(778/1 082) of the students had received knowledge about AIDS and 59.4% (643/1 082) of the students had received knowledge about sexual health in the past, from the three schools. There appeared statistical differences among the two types of schools. 7.0% (76/1 082) of the students reported having had sexual behaviors. Results from the multivariate logistic regression analysis showed that, in vocational senior middle school, the awareness rate of AIDS knowledge was lower (compared with general middle school, OR=0.41, 95%CI: 0.29-0.59) and in the following items, including no need in receiving safe sex health knowledge (compared with needs of safe sex health knowledge, OR=0.62, 95%CI: 0.43-0.91), no interests in reading AIDS-related health education materials (compared with attention to AIDS health education materials, OR=0.41, 95%CI: 0.30-0.56), not aware of safe sex behavior (compared with aware of safe sex behavior, OR=0.55, 95%CI: 0.39-0.77), not aware of correct condom use, compared with aware of correct condom use (OR=0.33, 95%CI: 0.24-0.46). Conclusions: The overall awareness rate of AIDS knowledge was low among the first-grade students in Tianjin. Some students reported of already having had insertive sex, sexual health knowledge has a significant impact on AIDS knowledge, suggesting that sexual health education and AIDS health education among high school students be strengthened in Tianjin.

[Effect of famine exposure on the risk of chronic disease in later life among population in Harbin].

Objective: To study the relations between famine exposure and the risk of chronic diseases as diabetes mellitus, obesity, hypertension, coronary heart disease and stroke in the population of Harbin. Methods: Our data was collected from the baseline survey-the China Kadoorie Biobank project (CKB) in Harbin. Retrospective cohort study design was used. Related risks on chronic diseases including diabetes mellitus, obesity, hypertension, coronary heart disease and stroke, were compared among the famine exposed or non-exposed people, respectively by logistic analysis method. Results: After adjusted for factors as age, sex, physical activity, smoking, alcohol intake, diet, family history of diseases, it appeared that the factor 'famine exposure' had increased the risks of diseases as obesity (OR=1.204, 95%CI: 1.104-1.313, P<0.01), hypertension (OR=1.315, 95%CI: 1.210-1.429, P<0.01) and coronary heart disease (OR=1.495, 95%CI: 1.369-1.632, P<0.01). The lower the age of population being exposed to famine, the greater the risk of the development of all kinds of chronic diseases. Conclusions: Famine exposure appeared a risk factor for obesity, hypertension, and coronary heart disease. It is of great significance to ensure the life-long nutrition of the people, especially in the early and adolescent stages, to prevent obesity, hypertension, and coronary heart disease in their later lives.

Aspirin Blocks Orthodontic Relapse via Inhibition of CD4+ T Lymphocytes.

Immunologic response plays an important role in orthodontic tooth movement (OTM) and relapse. Nonsteroidal anti-inflammatory drugs, such as aspirin, affect immune cells and clinical orthodontic treatment. However, the mechanisms by which nonsteroidal anti-inflammatory drugs regulate immune cells to affect orthodontic relapse are unclear. In this study, male Sprague-Dawley rats were grouped as relapse and relapse + aspirin for 10 d after 14 d of OTM. Silicone impressions of the rats' maxillary dentitions were obtained to record the distance of OTM at the indicated time point. CD4+ T lymphocytes in spleen were examined by flow cytometry. Serum levels of type 1 T-helper (Th1) cell-associated cytokines tumor necrosis factor α (TNF-α), and interferon γ (IFN-γ) were determined through enzyme-linked immunosorbent assay. The effects of aspirin on CD4+ T and Th1 cells were also analyzed in vitro. Aspirin treatment significantly reduced the relapse rate. More interestingly, injection of CD25 neutralizing antibody basiliximab or TNF-α inhibitor etanercept can significantly reduce the relapse rate as well. Correspondingly, aspirin treatment significantly accelerated the decrease of orthodontic force-induced secretion of TNF-α and IFN-γ in serum and the expression of TNF-α and IFN-γ in periodontal ligament during relapse. Furthermore, aspirin treatment in vitro significantly repressed the differentiation of CD4+ T and Th1 cells. Overall, results indicated that aspirin treatment can block orthodontic relapse by regulating Th1 cells.

Induction of unscheduled DNA synthesis in rat stomach mucosa by glandular stomach carcinogens.

Induction of unscheduled DNA synthesis (UDS) (repair DNA synthesis) in stomach pyloric mucosa of the F344/- DuCrj rat was examined in in vitro organ cultures in the presence of tritiated thymidine ([3H]dThd) and hydroxyurea after administration of chemical carcinogens in vivo. The DNA fraction was extracted from the cultured tissue, and the incorporation of [3H]dThd into DNA was determined in a liquid scintillation counter. DNA concentration was determined spectrophotometrically with either diphenylamine or 3,5- diaminobenzoic acid. A good correlation between induction of UDS and site specificity of carcinogens was observed. The glandular stomach carcinogens N-methyl-N'-nitro-N-nitrosoguanidine (CAS: 70-25-7), N-ethyl-N'-nitro-N-nitrosoguanidine (CAS: 63885-23-4), N-propyl-N'-nitro-N-nitrosoguanidine, 4-nitroquinoline 1-oxide (CAS: 56-57-5), and N-nitroso-N-methylurethane (CAS: 615-53-2) induced UDS in the pyloric mucosa of the stomach. UDS could be detected 2-4 hours after administration of carcinogens in vivo by the present method. The forestomach carcinogens 1-methyl-1-nitrosourea (CAS: 684-93-5) and aristolochic acid (CAS: 1398-06-7) and the nongastric carcinogens 2-acetylaminofluorene (CAS: 53-96-3), 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole, 3-amino-1-methyl-5H-pyrido[4,3-b]indole, and dimethylnitrosamine (CAS: 62-75-9) did not induce UDS in the pyloric mucosa.

Gastric- and intestinal-type properties of human gastric cancers transplanted into nude mice.

The gastric- and intestinal-type properties of 15 human gastric cancers, which were transplanted into nude mice, were studied biochemically and histologically. Enzyme activities were determined in the crude extracts of cancer tissues: pepsinogen isozymes as gastric marker enzymes; and sucrase, aminopeptidase (microsomal), and alkaline phosphatase as intestinal marker enzymes. By hematoxylin and eosin staining and paradoxical concanavalin A staining, gastric cancer tissues were classified into gastric type (pyloric gland cell type and surface mucous cell type) and intestinal type (goblet cell type and intestinal absorptive cell type). On the basis of their properties, human gastric cancers were classified into four types: (a) intestinal type; (b) gastric type; (c) intestinal plus gastric type; and (d) unclassified type, showing no gastric- or intestinal-type properties. Of six well-differentiated adenocarcinomas, four were of intestinal type, one of gastric type, and one of intestinal plus gastric type. All of the intestinal-type carcinomas showed sucrase activity. Of the three signet ring cell carcinomas, one was classified as a gastric type, one as an intestinal plus gastric type, and one as an unclassified type. Of the six poorly differentiated adenocarcinomas, five were of the intestinal type and one of the unclassified type. The present results clearly showed the appearance of intestinal-type properties in gastric cancer cells not only in so-called intestinal-type carcinomas, but also in diffuse-type carcinomas.

[Mortality rate and influencing factors among index partners of HIV discordant couples in the context of antiretroviral therapy, Zhumadian, Henan province, 2008-2014].

OBJECTIVE: To analyze the mortality rate in index partners of HIV discordant couples in the context of antiretroviral therapy and related factors in Zhumadian, Henan province. METHODS: An open and prospective cohort survey was conducted annually during 2008-2014. Information about the index partners ' demographic characteristics, CD4(+) T(CD4)cell counts, viral load, AIDS-related syndromes, date of therapy initiation, level of treatment institution, and outcome of death were collected. Life table was used to calculate survival rate, and Cox proportional hazards model was used to analyze the mortality related factors. RESULTS: Among 4 196 index partners receiving antiretroviral therapy between 2008 and 2014, 350 were fatal, with the mortality rate of 1.88 per 100 person years(95%CI: 1.86-1.90). The survival rates of these patients with the duration of follow-up from 1 to 7 years were 99.64%, 97.70%, 95.62%, 93.54%, 91.25%, 86.86% and 82.36% respectively. Factors related to the survival of the index partners of HIV discordant couples were age ≥ 50 years(HR=3.48, 95% CI: 2.67-4.55), junior high school or above education level(HR=0.75, 95% CI: 0.56-1.00), baseline CD4 cell counts >350 cells per µl(HR=0.55, 95% CI: 0.42-0.72), baseline viral load >400 copies/ml(HR=1.71, 95% CI: 1.32-2.22)and initiating treatment prior to cohort entry(HR=0.65, 95% CI: 0.41-1.05). CONCLUSIONS: The mortality rate of index partners receiving antiretroviral therapy was relatively low, indicating a better effect of long-term treatment on patients' survival. It is necessary to further identify and control the key factors influencing the survival of patients receiving therapy, improve the timeliness and adherence of the treatment, keeping patients' immunity level. Meanwhile, much attention should be paid to the drug resistance due to long term treatment and treatment failure.

Evaluation of dry feeding and liquid feeding to lactating sows under high temperature environment.

BACKGROUND: Liquid feeding system has been introduced to domestic swine farms, but negative cognition about liquid feeding system has been remained for feed waste decay related with poor management and microbial contamination. For these reasons, this study was conducted to evaluate the effects of feeding method in lactating sows. METHODS: A total of 30 mixed-parity (average 4.13) lactating sows (Yorkshire × Landrace) with an initial BW of 218.8 ± 19.5kg was used in a 3 week trial. Sows were allotted to 1 of 2 treatments in a completely randomized design by their body weight, backfat thickness, parity and alive litter weight. One of treatments was dry feeding and the other was liquid feeding (water to feed ratio, 1:1). Experimental diets contained 3265 kcal ME/kg, 12.6 % CP, 5.76 % EE, 1.09 % total lysine, 0.25 % total methionine, as fed basis. RESULTS: Dry feeding treatment had high body weight loss rather than liquid feeding treatment (P = 0.04). Dry feeding treatment had tendency to increase litter weight at 21d of lactation (P = 0.06) and litter weight gain (P = 0.04) during lactation period (0-3 week). Sows fed dry feeding method made milk containing high content of casein and total solid rather than sows fed liquid feeding method (P = 0.04). In addition, dry feeding treatment had tendency to higher content of milk fat, protein and solid not fat on 21d of lactation (P = 0.07). Sows fed dry feeding type also showed higher milk energy content in milk of 21d lactation (P = 0.05). Furthermore, liquid feeding treatment showed high occurrence in feed waste during lactation period (P < 0.01). CONCLUSION: Dry feeding method was more suitable feeding method to lactating sows under high temperature environment like lactating barn.

Effect of X-ray irradiation on pepsinogen and on induction of intestinal-type crypts in the pyloric mucosa of the Wistar rat stomach.

The effects of X-ray irradiation on the content and isozyme pattern of pepsinogen and on the induction of intestinal-type crypts in the pyloric mucosa of rat stomach were compared with the previously reported effects of chemical carcinogens, N-methyl-N'-nitro-N-nitrosoguanidine, N-ethyl-N'-nitro-N-nitrosoguanidine and N-propyl-N'-nitro-N-nitrosoguanidine. The gastric regions of 6-week-old male Wistar rats were irradiated with X-rays (1,000 rad) twice at an interval of 3 days. A decrease in pepsinogen content in the pyloric mucosa was observed between 2 and 52 weeks after the irradiation, as had been found in the previous experiments using chemical stomach carcinogens. However, in contrast to the latter experiment, in which the isozyme pattern was altered, the pepsinogen isozyme pattern did not change throughout the observation period after irradiation. Morphologically, intestinal-type crypts appeared and increased gradually after irradiation as after the administration of chemical stomach carcinogens. However, Paneth cells were either not observed or only rarely observed in rats treated with carcinogens, whereas they were found in crypts in the stomachs of the irradiated rats.