Associations of combined healthy lifestyles with cancer morbidity and mortality among individuals with diabetes: results from five cohort studies in the USA, the UK and China.

AIMS/HYPOTHESIS: Cancer has contributed to an increasing proportion of diabetes-related deaths, while lifestyle management is the cornerstone of both diabetes care and cancer prevention. We aimed to evaluate the associations of combined healthy lifestyles with total and site-specific cancer risks among individuals with diabetes. METHODS: We included 92,239 individuals with diabetes but without cancer at baseline from five population-based cohorts in the USA (National Health and Nutrition Examination Survey and National Institutes of Health [NIH]-AARP Diet and Health Study), the UK (UK Biobank study) and China (Dongfeng-Tongji cohort and Kailuan study). Healthy lifestyle scores (range 0-5) were constructed based on current nonsmoking, low-to-moderate alcohol drinking, adequate physical activity, healthy diet and optimal bodyweight. Cox regressions were used to calculate HRs for cancer morbidity and mortality, adjusting for sociodemographic, medical and diabetes-related factors. RESULTS: During 376,354 person-years of follow-up from UK Biobank and the two Chinese cohorts, 3229 incident cancer cases were documented, and 6682 cancer deaths were documented during 1,089,987 person-years of follow-up in the five cohorts. The pooled multivariable-adjusted HRs (95% CIs) comparing participants with 4-5 vs 0-1 healthy lifestyle factors were 0.73 (0.61, 0.88) for incident cancer and 0.55 (0.46, 0.67) for cancer mortality, and ranged between 0.41 and 0.63 for oesophagus, lung, liver, colorectum, breast and kidney cancers. Findings remained consistent across different cohorts and subgroups. CONCLUSIONS/INTERPRETATION: This international cohort study found that adherence to combined healthy lifestyles was associated with lower risks of total cancer morbidity and mortality as well as several subtypes (oesophagus, lung, liver, colorectum, breast and kidney cancers) among individuals with diabetes.

A basic/helix-loop-helix transcription factor controls leaf shape by regulating auxin signaling in apple.

Climate-driven phenological change across local spatial gradients leads to leaf shape variation. At higher elevations, leaves of broadleaf species tend to become narrower, but the underlying molecular mechanism is largely unknown. In this study, a series of morphometric analyses and biochemical assays, combined with functional identification in apple, were performed. We show that the decrease in apple leaf width with increasing altitude is controlled by a basic/helix-loop-helix transcription factor (bHLH TF), MdbHLH3. The MdbHLH3-overexpressing lines have a lower transcript abundance of MdPIN1 encoding an auxin efflux carrier but a higher transcript abundance of MdGH3-2 encoding a putative auxin amido conjugate synthase, resulting in a lower free auxin concentration; feeding the transgenic leaves with exogenous auxin partially restores leaf width. MdbHLH3 transcriptionally suppresses and activates MdPIN1 and MdGH3-2, respectively, by specifically binding to their promoters. This alters auxin homeostasis and transport, consequently leading to changes in leaf shape. These findings suggest that the bHLH TF MdbHLH3 directly modulates auxin signaling in controlling leaf shape in response to local spatial gradients in apple.

Transcriptional and physiological analyses of short-term Iron deficiency response in apple seedlings provide insight into the regulation involved in photosynthesis.

BACKGROUND: Iron (Fe) is an essential micronutrient for plants. Utilization of Fe deficiency-tolerant rootstock is an effective strategy to prevent Fe deficiency problems in fruit trees production. Malus halliana is an apple rootstock that is resistant to Fe deficiency; however, few molecular studies have been conducted on M. halliana. RESULTS: To evaluate short-term molecular response of M. halliana leaves under Fe deficiency condition, RNA sequencing (RNA-Seq) analyses were conducted at 0 (T1), 0.5 (T2) and 3 d (T3) after Fe-deficiency stress, and the timepoints were determined with a preliminary physiological experiment. In all, 6907, 5328, and 3593 differentially expressed genes (DEGs) were identified in pairs of T2 vs. T1, T3 vs. T1, and T3 vs. T2. Several of the enriched DEGs were related to heme binding, Fe ion binding, thylakoid membranes, photosystem II, photosynthesis-antenna protein, porphyrin and chlorophyll metabolism and carotenoid biosynthesis under Fe deficiency, which suggests that Fe deficiency mainly affects the photosynthesis of M. halliana. Additionally, we found that Fe deficiency induced significant down-regulation in genes involved in photosynthesis at T2 when seedlings were treated with Fe-deficient solution for 0.5 d, indicating that there was a rapid response of M. halliana to Fe deficiency. A strong up-regulation of photosynthesis genes was detected at T3, which suggested that M. halliana was able to recover photosynthesis after prolonged Fe starvation. A similar expression pattern was found in pigment regulation, including genes for coding chlorophyllide a oxygenase (CAO), ß-carotene hydroxylase (ß-OHase), zeaxanthin epoxidase (ZEP) and 9-cis-epoxycarotenoid dioxygenase (NCED). Our results suggest that pigment regulation plays an important role in the Fe deficiency response. In addition, we verified sixteen genes related to photosynthesis-antenna protein, porphyrin and chlorophyll metabolism and carotenoid biosynthesis pathways using quantitative real-time PCR (qRT-PCR) to ensure the accuracy of transcriptome data. Photosynthetic parameters, Chl fluorescence parameters and the activity of Chlase were also determined. CONCLUSIONS: This study broadly characterizes a molecular mechanism in which pigment and photosynthesis-related regulations play indispensable roles in the response of M. halliana to short-term Fe deficiency and provides a basis for future analyses of the key genes involved in the tolerance of Fe deficiency.

Transcriptome analysis in Malus halliana roots in response to iron deficiency reveals insight into sugar regulation.

Iron (Fe) deficiency is a frequent nutritional problem limiting apple production in calcareous soils. The utilization of rootstock that is resistant to Fe deficiency is an effective way to solve this problem. Malus halliana is an Fe deficiency-tolerant rootstock; however, few molecular studies have been conducted on M. halliana. In the present work, a transcriptome analysis was combined with qRT-PCR and sugar measurements to investigate Fe deficiency responses in M. halliana roots at 0 h (T1), 12 h (T2) and 72 h (T3) after Fe deficiency stress. Total of 2473, 661, and 776 differentially expressed genes (DEGs) were identified in the pairs of T2 vs. T1, T3 vs. T1, and T3 vs. T2, respectively. Several DEGs were enriched in the photosynthesis, glycolysis and gluconeogenesis, tyrosine metabolism and fatty acid degradation pathways. The glycolysis and photosynthesis pathways were upregulated under Fe deficiency. In this experiment, sucrose accumulated in Fe-deficient roots and leaves. However, the glucose content significantly decreased in the roots, while the fructose content significantly decreased in the leaves. Additionally, 15 genes related to glycolysis and sugar synthesis and sugar transport were selected to validate the accuracy of the transcriptome data by qRT-PCR. Overall, these results indicated that sugar synthesis and metabolism in the roots were affected by Fe deficiency. Sugar regulation is a way by which M. halliana responds to Fe deficiency stress.

Association of Combined Healthy Lifestyles With Cardiovascular Disease and Mortality of Patients With Diabetes: An International Multicohort Study.

OBJECTIVE: To prospectively examine the associations of combined lifestyle factors with incident cardiovascular disease (CVD) and mortality in patients with diabetes. PATIENTS AND METHODS: Patients with prevalent diabetes were included from 5 prospective, population-based cohorts in China (Dongfeng-Tongji cohort and Kailuan study), the United Kingdom (UK Biobank study), and the United States (National Health and Nutrition Examination Survey and National Institutes of Health-AARP Diet and Health Study). Healthy lifestyle scores were constructed according to non-current smoking, low to moderate alcohol drinking, regular physical activity, healthy diet, and optimal body weight; the healthy level of each lifestyle factor was assigned 1 point, or 0 for otherwise, and the range of the score was 0 to 5. Cox proportional hazards models were used to estimate hazard ratios for incident CVD, CVD mortality, and all-cause mortality adjusting for sociodemographic, medical, and diabetes-related factors, and outcomes were obtained by linkage to medical records and death registries. Data were collected from October 18, 1988, to September 30, 2020. RESULTS: A total of 6945 incident CVD cases were documented in 41,350 participants without CVD at baseline from the 2 Chinese cohorts and the UK Biobank during 389,330 person-years of follow-up, and 40,353 deaths were documented in 101,219 participants from all 5 cohorts during 1,238,391 person-years of follow-up. Adjusted hazard ratios (95% CIs) comparing patients with 4 or 5 vs 0 or 1 healthy lifestyle factors were 0.67 (0.60 to 0.74) for incident CVD, 0.58 (0.50 to 0.68) for CVD mortality, and 0.60 (0.53 to 0.68) for all-cause mortality. Findings remained consistent across different cohorts, subgroups, and sensitivity analyses. CONCLUSION: The international analyses document that adherence to multicomponent healthy lifestyles is associated with lower risk of CVD and premature death of patients with diabetes.

Sanguinarine Attenuates Neuropathic Pain in a Rat Model of Chronic Constriction Injury.

OBJECTIVE: There is still no effective treatment of neuropathic pain. Sanguinarine is a natural plant medicine with anti-inflammatory effects, but its effect on neuropathic pain remains unclear. This study was aimed at investigating the potential of sanguinarine to attenuate neuropathic pain. METHODS: Neuropathic pain was induced by chronic constriction injury (CCI) of the sciatic nerve. Rats were randomly divided into several groups: sham, CCI, CCI+SG (1.00 mg/kg), CCI+SG (2.50 mg/kg), and CCI+SG (6.25 mg/kg). SG was injected intraperitoneally from the day of surgery every three days. The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were recorded before surgery and on days 1, 3, 7, and 14 after surgery. The microglia in the spinal dorsal horn were examined by immunofluorescence. p38 MAPK expression in the spinal dorsal horn was detected by PCR and Western blot analysis. Cytokine levels in the spinal dorsal horn were measured by ELISA. RESULTS: MWT and TWL were significantly reduced in the CCI group, but sanguinarine recovered MWT and TWL in the CCI group. In addition, sanguinarine inhibited the activation of microglia and decreased the expression of p-p38 and TNF-α, IL-1ß, and IL-6 in the spinal dorsal horn of the CCI group in a dose-dependent manner. CONCLUSIONS: Our results suggest that sanguinarine can attenuate neuropathic pain via inhibiting the activation of microglia and the activation of the p38 MAPK signaling pathway.

[The distribution and influential factors of serum high sensitivity C-reactive protein in general population].

OBJECTIVE: To observe the distribution and influence factors of serum high sensitivity C-reactive protein (hs-CRP) in general population. METHODS: In a cross-sectional population survey, a total of 101 510 subjects who were employed by Kailuan Group had been carried out a healthy examination in the period of 2006 to 2007. In the statistical analysis, we observed 91 123 subjects (males 72 805, females 18 318) who had full information and met the inclusion criteria of the study. RESULTS: (1) The geometric means of hs-CRP were 0.70 mg/L, 0.70 mg/L and 0.73 mg/L in all subjects, males and females, respectively, the 95th percentiles were 6.28 mg/L, 6.20 mg/L and 6.49 mg/L, respectively. The concentrations of hs-CRP increased with age in both males and females (P trend = 0.001). Serum hs-CRP geometric mean was 0.54 mg/L and the 95th percentile was 5.40 mg/L in health group, while the geometric mean was 0.80 mg/L and the 95th percentile was 6.57 mg/L in non-health group. (2) Multiple linear regression analysis showed that concentrations of hs-CRP were positively associated with gender, age, systolic blood pressure, body mass index, total cholesterol, triglycerides, fasting blood glucose, smoking history, history of coronary heart disease and stroke history, but concentrations of hs-CRP were inversely related with diastolic blood pressure, high-density lipoprotein cholesterol and alcohol history. CONCLUSION: Serum concentrations of hs-CRP level increased with age, concentrations of hs-CRP were higher in females than males; a variety of cardiovascular factors effected the concentrations of hs-CRP.

[The risk predictive value of high sensitivity C-reactive protein level for new hemorrhagic stroke events].

OBJECTIVE: To study the risk prediction for new intracerebral hemorrhage (ICH) with high sensitivity C-reactive protein (hs-CRP) level. METHODS: In a retrospective, nested, case-controlled study, 323 cases of ICH were identified and matched with 646 controls. The hs-CRP levels at baseline were compared between the two groups. The relevance of different hs-CRP levels and the risk of ICH were analyzed. RESULTS: The ICH group had a higher median hs-CRP levels (1.10 mg/L) as compared with the control group (0.66 mg/L) with significant difference (P<0.01). In addition, the increase of risk associated with hs-CRP levels was primarily observed in the individuals with the highest quartile of hs-CRP levels (>2.12 mg/L). These patients had an increased risk of ICH (OR 2.58, 95%CI 1.77 to 3.76) as compared with those in the lowest quartile (≤0.30 mg/L). Individuals with baseline hs-CRP levels above the specified cut point of 3 mg/L or more and those in the 80th percentile were at a markedly increased risk of ICH (for specified cut point of 3 mg/L, OR 2.26, 95%CI 1.60 - 3.20, P<0.01; for 80th percentile, OR 2.24, 95%CI 1.60 - 3.13, P<0.01, respectively). CONCLUSIONS: Risk of ICH might be predicted with the level of hs-CRP. With the increase of hs-CRP level at baseline, the risk of ICH was increased.

Sanguinarine Attenuates Neuropathic Pain by Inhibiting P38 MAPK Activated Neuroinflammation in Rat Model.

BACKGROUND: Neuropathic pain seriously affects life quality, and it is urgent to develop novel drugs with high efficacy and few side effects. Sanguinarine (SG) is a natural plant medicine with anti-inflammatory and neuroprotection effects. This study aimed to investigate the effect of SG on chronic constriction injury (CCI)-induced neuropathic pain. MATERIALS AND METHODS: CCI rat model was established and rats were randomly divided into sham group, sham + SG group (6.25 mg/kg), CCI group, CCI + SG group (1.00, 2.50 and 6.25 mg/kg). The mechanical sensitivity and heat hypersensitivity of rats were monitored at different time points. Immunohistochemical, PCR, Western blot and ELISA were used to analyze p-p38 MAPK, NF-κB p65, TNF-α, IL-1ß, and IL-6 levels. RESULTS: The mechanical sensitivity and heat hypersensitivity significantly reduced in rats of CCI group, but significantly increased in rats of CCI+SG group. TNF-α, IL-1ß, and IL-6 levels significantly increased in the spinal cord of CCI rats, but significantly decreased in rats of CCI+SG group. In addition, p38 MAPK activator antagonized beneficial effects of SG on neuropathic pain. Overexpression of p38 MAPK reduced the mechanical sensitivity and heat hypersensitivity, and enhanced NF-κB activity and the expression of inflammatory factors in CCI rats. CONCLUSION: SG alleviates neuropathic pain via suppressing p38MAPK signaling and downregulating the expression of TNF-α, IL-1ß, IL-6 and NF-κB activation. SG may be a potential therapeutic agent to treat neuropathic pain.

Integrated physiologic, proteomic, and metabolomic analyses of Malus halliana adaptation to saline-alkali stress.

Saline-alkali stress is a severely adverse abiotic stress limiting plant growth. Malus halliana Koehne is an apple rootstock that is tolerant to saline-alkali stress. To understand the molecular mechanisms underlying the tolerance of M. halliana to saline-alkali stress, an integrated metabolomic and proteomic approach was used to analyze the plant pathways involved in the stress response of the plant and its regulatory mechanisms. A total of 179 differentially expressed proteins (DEPs) and 140 differentially expressed metabolites (DEMs) were identified. We found that two metabolite-related enzymes (PPD and PAO) were associated with senescence and involved in porphyrin and chlorophyll metabolism; six photosynthesis proteins (PSAH2, PSAK, PSBO2, PSBP1, and PSBQ2) were significantly upregulated, especially PSBO2, and could act as regulators of photosystem II (PSII) repair. Sucrose, acting as a signaling molecule, directly mediated the accumulation of D-phenylalanine, tryptophan, and alkaloid (vindoline and ecgonine) and the expression of proteins related to aspartate and glutamate (ASP3, ASN1, NIT4, and GLN1-1). These responses play a central role in maintaining osmotic balance and removing reactive oxygen species (ROS). In addition, sucrose signaling induced flavonoid biosynthesis by activating the expression of CYP75B1 to regulate the homeostasis of ROS and promoted auxin signaling by activating the expression of T31B5_170 to enhance the resistance of M. halliana to saline-alkali stress. The decrease in peroxidase superfamily protein (PER) and ALDH2C4 during lignin synthesis further triggered a plant saline-alkali response. Overall, this study provides an important starting point for improving saline-alkali tolerance in M. halliana via genetic engineering.

Association of Endometriosis-Associated Genetic Polymorphisms From Genome-Wide Association Studies With Ovarian Endometriosis in a Chinese Population.

Endometriosis is a common multifactorial disease caused by an interaction between multiple gene loci and environment. Four genome-wide association studies (GWASs) of endometriosis have identified several single-nucleotide polymorphisms (SNPs) associated with endometriosis. However, results from independent replication studies with different populations are inconsistent. The present study aims to evaluate whether the GWAS-derived susceptibility loci are correlated with the risk of the development of ovarian endometriosis in North Chinese women. This case-control study comprised 580 patients with ovarian endometriosis and 606 matched control women. Three SNPs were selected for this association study including rs10965235 in CDKN2BAS, rs2235529 located in LINC00339- WNT4, and rs12700667 in an intergenic region on 7p15.2. The results show that the G/A genotype of rs12700667 can significantly increase the risk of developing ovarian endometriosis when compared with the G/G genotype (odds ratio [OR] = 1.57, 95% confidence interval [CI] = 1.23-2.00). Similarly, the carriers with A allele showed a higher risk of ovarian endometriosis than those with G allele (OR = 1.23, 95% CI = 1.12-1.68). The study suggests that the endometriosis-associated genetic polymorphisms (rs12700667) from GWAS be associated with the risk of developing ovarian endometriosis in North Chinese women.

Predictive Value of Cumulative Blood Pressure for All-Cause Mortality and Cardiovascular Events.

The predictive value of cumulative blood pressure (BP) on all-cause mortality and cardiovascular and cerebrovascular events (CCE) has hardly been studied. In this prospective cohort study including 52,385 participants from the Kailuan Group who attended three medical examinations and without CCE, the impact of cumulative systolic BP (cumSBP) and cumulative diastolic BP (cumDBP) on all-cause mortality and CCEs was investigated. For the study population, the mean (standard deviation) age was 48.82 (11.77) years of which 40,141 (76.6%) were male. The follow-up for all-cause mortality and CCEs was 3.96 (0.48) and 2.98 (0.41) years, respectively. Multivariate Cox proportional hazards regression analysis showed that for every 10 mm Hg·year increase in cumSBP and 5 mm Hg·year increase in cumDBP, the hazard ratio for all-cause mortality were 1.013 (1.006, 1.021) and 1.012 (1.006, 1.018); for CCEs, 1.018 (1.010, 1.027) and 1.017 (1.010, 1.024); for stroke, 1.021 (1.011, 1.031) and 1.018 (1.010, 1.026); and for MI, 1.013 (0.996, 1.030) and 1.015 (1.000, 1.029). Using natural spline function analysis, cumSBP and cumDBP showed a J-curve relationship with CCEs; and a U-curve relationship with stroke (ischemic stroke and hemorrhagic stroke). Therefore, increases in cumSBP and cumDBP were predictive for all-cause mortality, CCEs, and stroke.

The different interactions of Colletotrichum gloeosporioides with two strawberry varieties and the involvement of salicylic acid.

The disease symptoms recognized as 'Anthracnose' are caused by Colletotrichum spp. and lead to large-scale strawberry (Fragaria×ananassa Duchesne) losses worldwide in terms of both quality and production. Little is known regarding the mechanisms underlying the genetic variations in the strawberry-Colletotrichum spp. interaction. In this work, Colletotrichum gloeosporioides (C. gloeosporioides) infection was characterized in two varieties exhibiting different susceptibilities, and the involvement of salicylic acid (SA) was examined. Light microscopic observation showed that C. gloeosporioides conidia germinated earlier and faster on the leaf surface of the susceptible cultivar compared with the less-susceptible cultivar. Several PR genes were differentially expressed, with higher-amplitude changes observed in the less-susceptible cultivar. The less-susceptible cultivar contained a higher level of basal SA, and the SA levels increased rapidly upon infection, followed by a sharp decrease before the necrotrophic phase. External SA pretreatment reduced susceptibility and elevated the internal SA levels in both varieties, which were sharply reduced in the susceptible cultivar upon inoculation. The less-susceptible cultivar also displayed a more sensitive and marked increase in the transcripts of NB-LRR genes to C. gloeosporioides, and SA pretreatment differentially induced transcript accumulation in the two varieties during infection. Furthermore, SA directly inhibited the germination of C. gloeosporioides conidia; NB-LRR transcript accumulation in response to SA pretreatment was both dose- and cultivar-dependent. The results demonstrate that the less-susceptible cultivar showed reduced conidia germination. The contribution of SA might involve microbial isolate-specific sensitivity to SA, cultivar/tissue-specific SA homeostasis and signaling, and the sensitivity of R genes and the related defense network to SA and pathogens.

Laparoscopic vaginoplasty using a single peritoneal flap: 10 years of experience in the creation of a neovagina in patients with Mayer-Rokitansky-Küster-Hauser syndrome.

OBJECTIVE: To assess anatomical and functional outcomes of a novel laparoscopic vaginoplasty technique using a single peritoneal flap (SPF) in patients with Mayer-Rokitansky-Küster-Hauser (MRKH) syndrome. DESIGN: Prospective follow-up study. SETTING: University-based tertiary-care hospital. PATIENT(S): Patients with MRKH syndrome (n = 83) and randomly selected frequency-matched age-comparable healthy women serving as controls (n = 85). INTERVENTION(S): From March 2004 to March 2014, a total of 83 patients with MRKH syndrome underwent laparoscopic vaginoplasty using an SPF. MAIN OUTCOME MEASURE(S): Intraoperative parameters, postoperative parameters, and anatomical outcomes were recorded. The functional results of patients who became sexually active were assessed using the Female Sexual Function Index (FSFI) questionnaire and were compared with those of the controls. RESULT(S): Laparoscopic vaginoplasty using an SPF was successfully performed in all 83 patients, with no intraoperative complications. The mean operative time and intraoperative blood loss were, respectively, 71.2 ± 18.9 minutes and 88.5 ± 57.2 ml. The mean length and width of the neovagina at the 6-month follow-up examination were, respectively, 8.2 ± 0.8 cm and 3.0 ± 0.6 cm. Anatomical success was achieved in all patients. At 12 months after surgery, functional success, as assessed by the FSFI questionnaire, was achieved in 95.3% of patients. The FSFI scores did not differ significantly between patients with MRKH and healthy women in a control group. CONCLUSION(S): Laparoscopic vaginoplasty using an SPF may be a feasible and effective approach that has satisfactory anatomical and functional outcomes for patients with MRKH syndrome.

[Effects of different drip irrigation modes on root distribution of wine grape 'Cabernet Sauvignon' in desert area of Northwest China].

To study the effects of different drip irrigation modes on the wine grape root distribution is the basis of formulating fertilization, irrigation, and over-wintering management practices for wine grape. Taking the wine grape "Cabernet Sauvignon" as test material, this paper studied the effects of different water-saving irrigation modes (drip irrigation under straw mulching, drip irrigation under plastic mulching, double-tube drip irrigation, and single-tube drip irrigation) on the root distribution of wine grape in the desert area of Northwest China, with the conventional furrow irrigation as the control. The root system of the "Cabernet Sauvignon" was distributed from 0 to 70 cm vertically, and from 0 to 120 cm horizontally. With double-tube drip irrigation, the root amount was the largest (138.3 roots per unit profile), but the root vertical distribution scope was narrowed by 20 cm, as compared to the control. Drip irrigation with straw mulching increased the root amount significantly, and increased the root horizontal distribution scope by 9.1%, as compared to the control. No significant difference was observed in the root number and root horizontal distribution scope between the drip irrigation under plastic mulching and the control, but the root vertical distribution scope with the drip irrigation under plastic mulching decreased by 20 cm. Single-tube drip irrigation increased the root number significantly, but had lesser effects on the root vertical or horizontal distribution, as compared to the conventional irrigation. It was suggested that the drip irrigation under straw mulching could be the best water-saving practice for the wine grape "Cabernet Sauvignon" in the study area.

[Inhibitory effect of VEGF antisense phosphorothioate oligodeoxynucleotides on the growth of human salivary adenoid cystic carcinoma xenografts in nude mice].

PURPOSE: To investigate the inhibitory effect of VEGF antisense phosphorothioate oligodeoxynucleoiides on the growth of human salivary adenoid cystic carcinoma (SACC) xenografts in nude mice. METHODS: The VEGF-ASODN was synthesised artificially. After the model of human SACC xenografts in nude mice was established, they were random1y divided into three groups: antisense group, scrambled group and normal saline group. A control group without cancer was also established. Antisense(66 µg), scrambled sequence(66 µg) and normal saline(once every 3 days and 7 times in all) were injected in three experimental groups, respectively. Two days after therapy, the mice were sacrificed. Serums were used for detection of VEGF protein. All tumors were measured and weighted. The quantity of VEGF mRNA and protein and PLI, MVD was detected by hybridization in situ and immunohistochemistry. SPSS13.0 software package was used for statistical analysis. RESULTS: The VEGF-ASODN could suppress the expression of VEGF in human SACC xenografts in nude mice and reduce VEGF protein in serum of nude mice significantly. It cou1d also reduce the volume and weight of xenografts and could reduce the expression of VEGF mRNA and its protein, PCNA and CD34. CONCLUSION: By inhibiting the expression of VEGF, VEGF-ASODN can inhabit proliferation of human SACC xenografts in nude mice.

[The effect of VEGF-ASODN transfection on expression of VEGF and growth in tongue squamous cancer cell line Tca8113].

PURPOSE: To evaluate the effect of transfection of vascular endothelial growth factor antisense oligonucleotide (VEGF-ASODN) on expression of vascular endothelial growth factor(VEGF) and growth in tongue cancer cells. METHODS: The VEGF-ASODN was synthesized with phosphorothioic acid. After transfecting with VEGF-ASODN in tongue cancer cells Tca8113, the quantity of VEGFmRNA and protein was detected by hybridization in situ and immunocytochemistry, and the quantity of VEGF protein in supernatant was detected by ELISA. FCM and MTT methods were used to detect cellular apoptosis and the activity of cells, respectively. The effect of transfection on the growth of cells was evaluated by growth curve. SPSS12.0 software package was used for statistical analysis. RESULTS: The quantity of VEGF mRNA, protein levels of VEGF in the cells and in culture fluid decreased after transfection of VEGF-ASODN (P<0.01,P<0.05).The ratio of apoptosis increased, the activity of cells also decreased after transfection VEGF-ASODN (P<0.01). CONCLUSIONS: Transfection with VEGF-ASODN in Tca8113 cells can inhibit the expression of VEGF remarkably. It can enhance cellular apoptosis and suppress growth of cells. Supported by Research Fund from Science and Technology Bureau of Taian City (Grant No.2007-46).

[The level of PPTA mRNA and its proteins expression in trigeminal ganglia in rats with occlusal recovery].

PURPOSE: To study the level of substance P(SP) and preprotachykinin A(PPTA) mRNA in trigeminal ganglia(TG)in rats with occlusal recoveryìand to discuss the mechanism of temporomandibular disorders. METHODS: 48 Wistar male rats were randomly divided into 3 experimental groups and 3 control groupsì8 rats in each group. The right maxillary and mandibular molars in the experimental groups were ground to the gingival level without occlusal contact. The occlusal contact was recovered by stoping grinding the molars gradually. The section of trigeminal ganglia were used for immunohistological and in situ hybridization study. Light microscope and microscoic photo analytic software were employed to detect the percentage of SP and PPTA mRNA positive neurons in the frozen section of TGs in 48 rats. SPSS10.0 software package were used for statistical analysis. RESULTS: The percentage of SP positive neurons in TG with unilateral chew experimental group decreased significantly compared to the control group (P<0.01,P<0.05).The decreased extent in the non-chewing side was much higher than that in the chewing side(P<0.05).The percentage of PPTA mRNA positive neurons was significantly higher in both of the chewing and non-chewing sides of the unilateral chew experimental group than that in the control group (P<0.05,P<0.01)ì and that non-chewing side was significantly higher than that in the chewing side(P<0.01).There was no significant difference in the percentage of SP and PPTA mRNA positive neurons between the early occlusal reconstruction the experimental group and the control group(P>0.05), and there was no significant difference of those between the non-chewing side and the chewing side(P>0.05). The results of later occlusal recovery in the experimental group was same to that of the unilateral chew experimental group. CONCLUSIONS: The expressions of SP and PPTA mRNA in TG can recover normal in the early occlusal recovery but can not in the later occlusal recovery. SP and PPTA mRNA might participate in the histopathologic mechanism of temporomandibular disorders.

[Expressions of calcitonin gene -related peptide in rats with occlusal reconstruction].

PURPOSE: To investigate the expressions of calcitonin gene-related peptide (CGRP) in rats with occlusal reconstruction. METHODS: 30 Wistar male rats were randomly divided into 3 experimental groups and 3 control groups, 5 rats in each group. The molar of the right maxilla and mandible of rats in the experimental groups were ground to the gingival level without occlusal contact. The occlusal contact was recovered by stopping grounding molar of the rats in two of the experimental groups. The section of the disc and articular capsule Was subject to immunohistological study to evaluate the expressions of CGRP by using SABC method. Light microscope and microscopic photo analytic software were employed to detect calcitonin gene-related peptide-like immunoreactive (CGRP-LI) nerve fibers in frozen section of TMJs in 30 rats. SPSS10.0 software package were used for statistical analysis. RESULTS: The areas of CGRP-LI nerve fibers in unilateral chew group were significantly larger than those in the control group (P < 0.01, P < 0.05). The increment in the non-chewing side was significantly higher than that in the chewing side (P < 0.05). There was no significant difference in the areas of CGRP-LI nerve fibers between the early occlusal reconstruction group and the control group (P > 0.05) and there was no significant difference between the non-chewing side and the chewing side (P > 0.05). There was significant difference in the areas of CGRP-LI nerve fibers between the later occlusal reconstruction group and the control group (P < 0.01), while no significant difference was found between the non-chewing side and the chewing side (P > 0.05). CONCLUSION: The expressions of CGRP can recover normal in early occlusal reconstruction but not in later occlusal reconstruction.CGRP might participate in the histopathogenesis of temporomandibular joint disorders. Supported by Research Fund from Science and Technology Bureau of Taian City (Grant No.2003-52).