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ObjectiveTo clarify the relationship between intestinal flora and intestinal motility in rats with slow transit constipation （STC） and qi stagnation syndrome by conducting a pseudo-sterile experiment and fecal microbiota transplantation （FMT） technology. MethodsTwenty-four Wistar rats were randomly divided into normal group （n=6）， STC with qi stagnation pattern group （n=6） and pseudo-sterile group （n=12）. In the STC group with qi stagnation pattern， 3 mg/kg of loperamide suspension by intragastric administration combined with tail clamping stimulation were performed to establish the rat model of STC with qi stagnation pattern. After successful modeling， fresh feces from the rats in the STC with qi stagnation pattern group and the normal group were collected to prepare 100 mg/ml of fecal bacterial suspension. In the pseudo-sterile group， the antibiotic cocktail method was used （a mixed antibiotic suspension containing bacitracin， streptomycin sulfate， and neomycin sulfate at 20 mg/ml each was administered intragastrically） to establish pseudo-sterile rats model. After successful modeling， the rats were randomly divided into normal fecal bacterial liquid group and STC with qi stagnation pattern fecal bacterial liquid group， with six rats in each group， and then were given 10 ml/kg of the prepared corresponding rat fecal bacterial suspension by gavage. Rats in STC with qi stagnation pattern group were given an equal volume of sterile water by gavage. All groups were administered once a day for 7 consecutive days. The small intestinal propulsion rate of the STC with qi stagnation pattern group， the normal fecal bacterial liquid group， and STC with qi stagnation pattern fecal bacterial liquid group were compared. ELISA method was used to detect serum 5-hydroxytryptamine （5-HT） levels. Immunohistochemistry was used to detect the positive expression levels of 5-hydroxytryptamine 3 receptor （5-HT3R） and 5-hydroxytryptamine 4 receptor （5-HT4R） in colon tissue. Western blot method was used to detect the protein expression levels of tryptophan hydroxylase 1 （TPH1）， tryptophan hydroxylase 2 （TPH2）， serotonin transporter （SERT）， and monoamine oxidase A （MAO-A） in colon tissue. ResultsCompared to those in the normal fecal bacterial liquid group， the small intestinal propulsion rate， serum 5-HT level， positive expression of 5-HT3R and 5-HT4R in colon tissue， and protein expression of TPH1， TPH2， SERT and MAO-A significantly decreased in the STC with qi stagnation pattern group and STC with qi stagnation pattern fecal bacterial liquid group （P＜0.05）. There was no statistically significant difference in the indicators between the STC with qi stagnation pattern group and STC with qi stagnation pattern fecal bacterial liquid group （P＞0.05）. ConclusionThe intestinal flora in STC rats with qi stagnation pattern can lead to a slowdown in intestinal transmission function， whose mechanism may be related to intestinal motility disorders affected by the synthesis， transport， metabolism and other pathways of 5-HT.
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Tripterygium wilfordii is a valuable medicinal plant rich in biologically active diterpenoids, but there are few studies on the origins of these diterpenoids in its secondary metabolism. Here, we identified three regions containing tandemly duplicated diterpene synthase genes on chromosomes (Chr) 17 and 21 of T. wilfordii and obtained 11 diterpene synthases with different functions. We further revealed that these diterpene synthases underwent duplication and rearrangement at approximately 2.3-23.7 million years ago (MYA) by whole-genome triplication (WGT), transposon mediation, and tandem duplication, followed by functional divergence. We first demonstrated that four key amino acids in the sequences of TwCPS3, TwCPS5, and TwCPS6 were altered during evolution, leading to their functional divergence and the formation of diterpene secondary metabolites. Then, we demonstrated that the functional divergence of three TwKSLs was driven by mutations in two key amino acids. Finally, we discovered the mechanisms of evolution and pseudogenization of miltiradiene synthases in T. wilfordii and elucidated that the new function in TwMS1/2 from the terpene synthase (TPS)-b subfamily was caused by progressive changes in multiple amino acids after the WGT event. Our results provide key evidence for the formation of diverse diterpenoids during the evolution of secondary metabolites in T. wilfordii.
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Objective To investigate the effects of subthreshold micropulse yellow laser (577 nm) on vascular endothelial growth factor (VEGF),nerve growth factor (NGF) and Chemerin expressions in retina of early stage diabetic rats.Methods A total of 40 Brown Norway rats were treated with streptozocin (65 mg · kg-1) to establish the diabetic model.20 diabetic BN rats' right eyes were received subthreshold micropulse yellow laser (577 run) therapy after 2 weeks.The left eyes were used as control group.At 3 days,7 days,14 days,28 days after laser therapy,5 BN rats were randomly chosen to perform RT-PCR and Weston-blot.The expressions of mRNA and protein of VEGF,NGF and Chemerin were analyzed.Results The expression of VEGF mRNA and protein increased in control group at 3 days,7 days,14 days and 28 days (all P ＜ 0.05).Compared with the control group,VEGF mRNA and protein decreased in the subthreshold micropulse yellow laser (577 nm) group (all P ＜ 0.05).The expression of NGF mRNA and protein decreased in the control group at 3 days,7 days,14 days and 28 days (all P ＜ 0.05),however,the difference was not statistically significant between 3 days and 7 days(P ＞0.05).Compared with control group,NGF mRNA and protein increased in the subthreshold micropulse yellow laser (577 nm) group (all P ＜ 0.05),with maximum expression at 14 days.The expression of chemerin mRNA and protein increased at 3 days,7 days,14 days and 28 days in the control group (all P ＜0.05).Compared with the control group,chemerin mRNA and protein decreased in the subthreshold micropulse yellow laser (577 urn) group (all P ＜ 0.05).Conclusion Subthreshold micropulse yellow laser (577 urn) can suppress VEGF,Chemerin expression and upregulate NGF expression in early stage diabetic rats.
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Objective To explore the relationship of course and recurrence after norms antiviral treatment for patients with chronic hepatitis B. Methods All cases were confirmed by standard use of nucleos drug treatment,after treatment,followed up for 104 weeks; patients after stopping the treatment and contiued to conslidate 1 year, set the control group,patients after stopping the treatment and tiued to consolidate 3 years, as observation group,the recurrence rates and recurrence time were compared. Results The relapse rate of observation group was significantly lower than the control group(χ2=3.877,P<0.05). Control group of 38 cases,31 cases of recurrence after treatment,included 2 cases of recurrence within 2 weeks,25 cases of recurrence within 24 weeks; 4 cases relapsed after 24 weeks.Observation group of 30 cases,18 cases of relapse after discontinuation,1 case relapsed in two weeks,15 cases of recurrence within 24 weeks,2 cases relapsed after treatment 24 weeks. Conclusion The prolonged antiviral therapy can reduce the recur-rence rate of the time,but did not extend the time to recurrence,we recommend patients with chronic hepatitis B nu-cleoside (acid) drug treatment medication should be strictly in accordance with the relevant guidelines,emphasizing long-term medication to maintain efficacy,And standardize management with antiviral treatment.
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Objective To investigate the effects of leptin on the oxidative damage in human retinal pigment epithelial (RPE) cells.Methods Human RPE cells (ARPE-19) were cultured in vitro,and randomly divided into control group and insulin resistance group.RPE cells were treated with 0,10,100 ng/mL leptin for 24,48,72 hours respectively.Then the levels of reactive oxygen species (ROS) expression in RPE cells were detected by 2',7'-dichlorofluorescin-diacetate (DCFH-DA),and the levels of 8-hydroxy-2′-deoxyguanosine (8-OHdG) expression in RPE cells were observed by immunocytochemistry (ICC),and the levels of human 8-oxoguanine DNA glycosylase 1 (hOGG1) expression in lysate were measured by Western blot.Results After 24,48,72 hours,the level of ROS (Control group:F=37.136,37.178,49.634; P＜0.05.Insulin resistance group:F=9.822,28.881,71.150;P＜0.05),8-OHdG (Control group:F =88.643,390.920,1039.276; P ＜ 0.05.Insulin resistance group:F =273.311,299.155,82.237;P＜0.05) and hOGGl (Control group:F=470.062,1073.113,295.456;P＜0.05.Insulin resistance group:F =240.032,592.389,527.760 ; P＜0.05) expression increased significantly with the increase of leptin concentration in control group and insulin resistance group.Under the same leptin concentration,the level of 8-OHdG has a trend that it was higher in the insulin resistance group than the control group.After 24 hours,the difference of hOGGl expression between control group and insulin resistance group was not significant (F=23.392,P＞0.05).After 72 hours,the level of hOGGl expression was significantly higher in the insulin resistance group than the control group (F=129.394,P＜0.05).The level of hOGGl expression was significantly higher at 48 hours than that at 24 hours and 72 hours (P＜ 0.05).Conclusion Leptin could induce the oxidative damage of RPE cells in normal and insulin resistance status.With the increase of leptin concentration and time extended,the degree of oxidative damage and its repair were both increased.The degree of oxidative repair increased with the increase of leptin concentration,but decreased with time extended.
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Objective To study the relationship of the university students' procrastination and big five personality factors.Methods 461 university students were assessed by the simple edition of tuckman procrastination scale(TPS) and the big five personality inventory short form(NEO-FFI).Results TPS score of the student was (48.51 ± 7.66).There were 89.4% students respectively had different level of procrastination behavior.There were no significant differences in university students by the gender(47.95 ±7.84 vs 48.95 ± 7.43 ) ,whether one-child students(48.64 ± 7.49 vs 48.45 ± 7.81 ) and grades.There were significant positive correlation between procrastination and the personality factor of neuroticism( r=0.235, P＜0.01 ) and significant negative correlation with extraversion ( r=-0.180, P ＜ 0.01 ) and conscientiousness ( r =-0.198, P ＜ 0.01 ).The big five personality factors of neuroticism,extraversion and conscientiousness could explain the 14.0% variation of procrastination of university students.Conclusion Procrastination is a very common phenomenon in the university students.The students with higher neuroticism, lower extraversion and conscientiousness of big five personality have more procrastination behavior.