摘要
Objective To investigate the predictive value of serum Nesfatin-1 and 25-hydroxyvitamin D3[25(OH)D3]levels for the short-term prognosis of status epilepticus(SE)in children.Methods A total of 104 children with SE admitted to the hospital from March 2020 to March 2023 were enrolled in the study,and the clinical data of the children were collected.According to the Glasgow outcome Scale(GOS)score at dis-charge,the children were divided into a good prognosis group(equal to 5 points)and a poor prognosis group(＜5 points).Univariate analysis and multivariate Logistic regression were used to analyze whether serum Nesfatin-1 and 25(OH)D3 levels were risk factors for poor short-term prognosis in children with SE.The re-ceiver operating characteristic(ROC)curve was drawn to analyze the predictive value of serum Nesfatin-1 and 25(OH)D3 levels for the short-term poor prognosis in children with SE.Results At discharge,85 children[81.73％(85/104)]with a GOS score of 5 were included in the good prognosis group,and 19 children[8.27％(19/104)]with a GOS score of＜5 were included in the poor prognosis group.There was no significant differ-ence in gender,age,previous history of epilepsy,and seizure types between the two groups(P＞0.05).There were significant differences in the duration of SE,the time from medication to seizure cessation,electroenceph-alogram(EEG)results,head CT results,and serum Nesfatin-1 and 25(OH)D3 levels between the two groups(P＜0.05).Multivariate Logistic regression analysis showed that SE duration＞60 min,abnormal head CT results,serum Nesfatin-1 and 25(OH)D3 levels were independent risk factors for the short-term poor progno-sis of children with SE(OR=1.945,2.343,1.731,1.505;P＜0.05).The area under the ROC curve of serum Nesfatin-1 and 25(OH)D3 levels combined to predict poor short-term prognosis of children with SE was 0.840(95％CI:0.732-0.949),which was better than that of serum Nesfatin-1 and 25(OH)D3 levels alone[0.607(95％CI:0.453-0.761),0.742(95％CI:0.604-0.880)],respectively.Conclusion Serum Nesfatin-1 and 25(OH)D3 levels are risk factors for poor prognosis in children with SE,and the combination of them has high predictive value for poor prognosis in children with SE.
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Objective To explore the medication rules of Professor ZHONG Guang-Ling's prescriptions for the treatment of lumbar disc herniation(LDH)based on data mining method,so as to provide reference for the treatment of LDH with Chinese medicine.Methods The prescriptions for the effective cases of outpatients of LDH treated by Professor ZHONG Guang-Ling in the recent 5 years were collected.The medication frequency of Chinese medicines in the included prescriptions and the distribution of their properties,flavors and meridian tropism were investigated.Moreover,the association rule analysis and cluster analysis of the high-frequency drugs were carried out.Results A total of 164 prescriptions were included and 168 Chinese medicines were used.The top 10 high-frequency drugs in descending order were Achyranthis Bidentatae Radix,Chuanxiong Rhizoma,Pheretima,Glycyrrhizae Radix et Rhizoma Praeparata cum Melle,Gentianae Macrophyllae Radix,Angelicae Sinensis Radix,Rehmanniae Radix Praeparata,Persicae Semen,Cyperi Rhizoma,and Carthami Flos.The properties of the prescribed drug were mainly warm and mild in nature,and bitter and pungent in flavor,and mainly had the meridian tropism of the liver,kidney and spleen meridians.According to the therapeutic actions,the drugs were mainly categorized as deficiency-supplementing drugs,dampness-removing and collateral-unblocking drugs,and blood-activating and stasis-removing drugs.The results of association rule analysis yielded 10 drug pairs,and cluster analysis yielded 6 core drug combinations.Conclusion For the treatment of LDH,Professor ZHONG Guang-Ling usually adopts the Chinese medicine for supplementing the deficiency and supporting healthy-qi,together with the medicines for nourishing the liver and kidney and regulating the spleen and stomach from the perspective of liver,kidney and spleen.Moreover,therapy of activating blood and removing stasis is also stressed,pathogen-eliminating medicines for removing dampness,unblocking collaterals and clearing heat are used based on syndrome differentiation,and then simultaneous application of purging and nourishing therapeutics is achieved through the utilization of purging method after supplementing method.
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Objective:To explore the clinical application and triage management value of using blood circulating cell-free DNA (cfDNA) (cysteine dioxygenase type 1 gene, CDO1, and Homeobox protein A9 gene, HOXA9) hypermethylation level to detect and diagnose ovarian cancer.Methods:A case-control study was conducted on patients who went for surgery at Chengdu Womens and Childrens Central Hospital from November 2022 to October 2023. Blood samples were collected before surgery for evaluation of cancer antigen 125 (CA125), human epididymis protein 4 (HE4), risk of ovarian malignancy algorithm (ROMA) score, and DNA methylation testing. The basic clinical information, biomarkers, and transvaginal ultrasound (TVS) information were collected simultaneously. Information from a total of 151 patients was collected, including 122 cases with benign pathology and 29 ovarian cancer cases. The pathologic diagnosis of ovarian tissue was defined as the gold standard. The multivariate logistic regression analysis was used to identify high-risk factors for ovarian cancer. The clinical efficacy of DNA methylation detection for ovarian cancer was analyzed using the area under curve (AUC).Results:The results showed that the age, menopausal status, CA125 and HE4 detection, ROMA score, positivity rate of CDO1 gene and HOXA9 gene single or combined testing in ovarian cancer patients were higher than those in the benign group and showed significant differences ( P<0.05). Among these detection protocols, the AUC of CDO1 and HOXA9 dual gene methylation testing for ovarian cancer was the highest at 0.936 (95% CI, 0.878-0.994), with 89.7% (95% CI 73.6%-96.4%) sensitivity and 97.5% (95% CI 93.0%-99.2%) specificity, respectively. The positive detection rate of CDO1 and HOXA9 dual gene methylation in early ovarian cancer FOGO I-II stage is 12/14 higher than other tests. Conclusion:Blood cfDNA methylation detection, a simple, non-invasive, and highly sensitive detection method, is superior to the current ovarian cancer testing in the risk assessment and early detection.
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Objective To explore the value of baseline plasma soluble type 2 myeloid cell trigger receptors(sTREM2)in evaluation for early hematoma enlargement in elderly patients with spontan-eous cerebral hemorrhage(SCH).Methods Clinical data of 240 patients with acute SCH admitted to our hospital from January 2020 to August 2022 were collected and analyzed retrospectively.According to the expansion of the hematoma volume,they were divided into non-expansion group(172 cases)and expanded group(68 cases).Baseline head CT scanning was performed in all patients within 24 h of onset,clinical and imaging data were analyzed,and the volume of cerebral hematoma was calculated.Blood samples were collected immediately after admission and sTREM2 and galectin-3 levels were measured.Results Compared with the non-expansion group,the ex-panded group had larger cerebral hematoma volume,and increased levels of sTREM2,galectin-3,hs-CRP and TNF-a at admission(P＜0.01).Pearson correlation analysis showed that the expres-sion levels of sTREM2 and galectin-3 were positively correlated with cerebral hematoma at ad-mission(r=0.784,P=0.012;r=0.815,P=0.004).ROC curve analysis indicated that the sensi-tivity of combined serum sTREM2 and galectin-3 levels was significantly higher than that of sin-gle detection(85.59％vs 73.73％and 64.41％,P＜0.05),and the AUC value was 0.896(95％CI:0.741-0.932).Conclusion The baseline plasma level of sTREM2 is significantly increased in eld-erly SCH patients after early hematoma expansion.So,sTREM2 can be used as a risk marker for early expansion of hematoma,and its combined detection with galectin-3 shows higher diagnostic value.
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AIM To explore the effects of Zishui Qinggan Decoction on the mouse model of depression induced by chronic restraint stress(CRS)via ERK/GSK3β/CREB/BDNF signaling pathway.METHODS Except for those of the blank group,the mice of other groups were induced into depression models by CRS,and divided into the model group,the fluoxetine hydrochloride group(10 mg/kg)and the low,medium and high dose Zishui Qinggan Decoction groups(8.835,17.670 and 35.340 g/kg)for the corresponding drug intervention and simultanous CRS treatment.The mice had their sugar water preference experiment and behavior experiment on the 7th and 14th day after administration;the observation of the hippocampal morphological changes by HE staining,the detection of the superoxide dismutase(SOD)activity and malondialdehyde(MDA)level in serum by kits,the detection of levels of serum 5-hydroxytryptamine(5-HT),tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)by ELISA,the detection of the hippocampal mRNA expressions of BDNF,TNF-α and IL-1β by RT-qPCR method,and the detection of the hippocampal protein expressions of ERK1/2,p-ERK1/2,GSK3β,p-GSK3β,CREB and BDNF by Western blot method 14 days after administration.RESULTS Compared with the model group,after 14 days of administration,both fluoxetine hydrochloride group and medium-dose Zishui Qinggan Decoction group displayed increased preference rate of sugar water(P<0.01),shortened immobility time of tail suspension and forced swimming(P<0.01),improved hippocampal damage of nerve cells,decreased levels of serum MDA,TNF-α and IL-1β(P<0.05,P<0.01),increased SOD activity and 5-HT level(P<0.05,P<0.01),decreased hippocampal mRNA expressions of TNF-α and IL-1β(P<0.01),and decreased expressions of BDNF mRNA and p-ERK1/2,p-GSK3β,CREB and BDNF proteins(P<0.05,P<0.01).CONCLUSION Zishui Qinggan Decoction can improve the depression-like behaviors in mice exposed to CRS,and its mechanism may be related to the regulation of hippocampal ERK/GSK3β/CREB/BDNF signaling pathway.
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Objective:To investigate the molecular mechanism for regulation of trophoblast invasion by piR-3127964, which is differentially expressed in placental tissues of preeclamptic and healthy pregnant women.Methods:Placenta samples of healthy (control group, n=12) and preeclamptic pregnant (PE group, n=10) women who delivered by caesarean section and chorionic villi specimens of patients undergoing artificial abortion were collected in the Department of Obstetrics of the First Affiliated Hospital of Chongqing Medical University during November 2020 to August 2021. Total RNA was extracted from placenta samples and sequenced and the expression of piR-3127964 in different tissues was determined by real-time quantitative-polymerase chain reaction (qRT-PCR). The expressions of PIWI proteins including PIWIL-1, PIWIL-2 and PIWIL-3 in different tissues were detected by Western blot. The expressions of two candidate targets, guanine nucleotide-binding protein-like 3-like (GNL3L) mRNA and sialophorin (SPN) mRNA were evaluated by qRT-PCR after exogenous treating HTR-8/SVneo cells with mimics, inhibitor or negative control of piR-3127964, respectively. qRT-PCR was also used to detect the relative expression of GNL3L and SPN at mRNA level in placentas of all women. The interactions between GNL3L/SPN and piR-3127964 were analyzed by double luciferase reporter gene detection. The localization of piR-3127964 and SPN in chorionic villi was detected by fluorescence in situ hybridization and immunofluorescence. Transwell assay was performed to analyze the influence of piR-3127964 on the invasion of HTR-8/SVneo cells and the possible mechanism. Independent sample t-test, analysis of variance, and LSD post test were used for analysis Results:(1) Enrichment pathways of candidate targets predicted by differentially expressed piR-3127964 were associated with cell motility. There were statistically significant differences in piR-3127964 expression in villi, healthy and preeclamptic placentas (2.950±0.853 vs 1.036±0.303 vs 0.254±0.155, F=27.35, P<0.05), and piR-3127964 was predominantly expressed in extravillous cytotrophoblasts (EVTs). (2) The expression of PIWIL-3 protein in placentas of preeclamptic patients was significantly lower than that in healthy placentas and villi (0.810±0.400 vs 3.175±0.429 and 6.843±1.379, F=49.36, P<0.05). (3) Compared with the control group, exogenous piR-3127964 mimics (piR-mimics) and inhibitors (piR-inhibitor) significantly affected the expression of SPN mRNA (0.971±0.045 vs 0.732±0.010, F=6.50; 1.076±0.073 vs 1.293±0.092, F=7.58; both P<0.05), while the expression of GNL3L mRNA had no significant correlation with piR-3127964 level. (4) The luciferase activity of wild-type SPN (SPN-WT) plasmids was significantly affected by piR-mimics (1.010±0.049 vs 0.645±0.047, t=9.34, P<0.05) and piR-inhibitor (1.035±0.058 vs 1.397±0.015, t=－10.60, P<0.05). (5) SPN mRNA was significantly upregulated in placentas of preeclamptic patients than in healthy placentas (2.097±0.239 vs 1.305±0.290, t=－4.22, P<0.05), but no significant difference in the expression of GNL3L mRNA was observed. Immunofluorescence experiment showed that SPN was expressed in EVTs. (6) The invasive potential of HTR8/SVneo cells treated with piR-inhibitor was significantly inhibited, but this effect could be reversed by SPN knockdown (160.714±53.860 vs 371.667±103.061 and 344.333±120.267, F=9.76, both P<0.05). Conclusions:piR-3127964 expression is abnormally downregulated in placentas of preeclamptic patients, resulting in inhibition of trophoblasts invasion through upregulation of SPN expression, which may be related to the pathogenesis of preeclampsia.
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@#[摘 要] 目的：探讨乳腺癌特异基因1（BCSG1）与Hsa-circ-0026352在浸润性乳腺癌（IBC）遗传易感性中的交互作用。方法：选取2019年6月至2022年5月间武汉市中西医结合医院收治的100例IBC患者作为研究对象，采用免疫组化法检测IBC组织及其相应癌旁组织中BCSG1的表达，将研究对象按照IBC组织中BCSG1蛋白表达的高低分为阴性、弱阳性和强阳性组，统计三组患者的临床病理特征及雌激素受体（ER）、孕激素受体（PR）、表皮生长因子受体-2（HER2）、Hsa-circ-0026352的表达情况，采用Logistic回归方程和最大似然法分析BCSG1表达与上述参数的趋势性和交互作用。结果：与癌旁组织比较，IBC组织中BCSG1蛋白呈高表达（P<0.05）；BCSG1蛋白强阳性表达与淋巴结转移、分化程度、临床分期、HER2表达、 Hsa-circ-0026352表达有关联（P<0.05）；BCSG1强阳性表达与IBC存在交互作用（P<0.05）；BCSG1表达与IBC的交互作用在Hsa-circ-0026352阳性表达中最为显著（趋势P<0.001）；BCSG1表达与IBC的交互作用在临床分期Ⅲ期、低分化程度中最为显著（趋势P<0.001）。结论：BCSG1与IBC患病密切相关，且与Hsa-circ-0026352、临床分期、分化程度存在交互作用，可共同增加IBC患病风险性。
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Objective@#To analyze the influencing factors and physical and mental development of preschool children with iron deficiency anemia in Dongguan, so as to provide a reference for the prevention of iron deficiency anemia among preschool children.@*Methods@#A total of 118 preschool children with iron deficiency anemia who were examined in Dongguan Maternal and Child Health Center from January 2022 to December 2022 were enrolled in the anemia group, and 118 preschool healthy children who were examined in the hospital at the same time were enrolled in the control group. The physical and mental development of the children were evalucded in both groups. Demographic information and household per capita income were collected. The relationship between risk factors and iron deficiency anemia was analyzed by univariate analysis and multiple Logistic regression.@*Results@#The scores of fine motor skills, gross motor skills, adaptability, social communication, language ability and developmental quotient of children in anemia group were significantly lower than those in control group ( t =4.14, 5.46, 5.60, 5.50, 4.90, 5.83, P <0.01). The difference in scores of adaptability, fine motor skills, gross motor skills language ability, social communication and developmental quotient between the two groups increased with age ( F =390.56, 414.63, 437.35, 409.68, 407.20, 404.54, P < 0.05 ). Multivariate Logistic regression analysis showed that household income, history of past digestive disease, gestational age, maternal anemia during pregnancy, maternal education, consumption of meat, eggs and milk, and intake of nuts were all associated with iron deficiency anemia among preschool children in Dongguan ( OR =2.23，2.99，3.99，3.56，3.11，1.68，1.61, P < 0.05 ).@*Conclusion@#The physical and mental development of preschool children with iron deficiency anemia in Dongguan is slower than that of non anemia children of the same age, and the development delay becomes more obvious with increasing age. Attention should be paid to the prevention of iron deficiency anemia among preschool children. It is important to provide reasonable dietary guidance for children with high risk factors such as digestive disease history and prematurity.
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Cardiovascular disease (CVD) is the leading cause of mortality and healthy life expectancy loss, ranking first in causing the global burden of disease. In addition to the traditional CVD risk factors, such as hypertension and diabetes, environmental chemical pollutants may also play a role in the development of CVD. This paper summarizes the evidence regarding the relation of exposures to metal or metalloid and persistent organic pollutants with risk for CVD and introduces the research progress in the relation between the exposures to two environmental chemical pollutants and CVD risk. The study aims to provide scientific evidence for the effective prevention of CVD through the management of chemical pollutants in environment.
Subject(s)
Humans , Cardiovascular Diseases , Persistent Organic Pollutants , Metalloids , Hypertension , Environmental Pollutants摘要
Objective To identify the retention time of semen(stains)inside and outside the cadaver using Fourier Transform Infrared(FTIR)spectroscopy and chemometrics.Methods A simulated environment inside the porcine vagina was used to retain fresh semen for 0～168 hours inside and outside the body.Infrared spectroscopy data were collected,and chemometric methods including Principal Components Analysis(PCA)and Partial Least Squares(PLS)regression models were established.Results The infrared spectrum peak positions of semen(stains)inside and outside the body did not show significant changes over time.The PCA results showed good clustering relationships,and the predicted deviation value in the PLS regression model was 15.786 3 hours,with an R2 value of 0.922 4.The characteristic wavenumbers selected by the CARS algorithm were in the wavelength ranges of 1 000～1 180 cm-1,1 430～1 480 cm-1,and 1 700～1 760 cm-1.Conclusion Fourier Transform Infrared spectroscopy has the potential to identify the retention time of semen(stains)inside and outside the body,and there are differences in the degradation trends of semen(stains)inside and outside the body.
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OBJECTIVE@#To observe the effects of electroacupuncture (EA) on NLRP3 inflammasome and its downstream protein gastermin D (GSDMD) in rats with primary dysmenorrhea (PDM), and to explore the potential mechanism of EA on the treatment of PDM.@*METHODS@#Forty healthy female SD rats without pregnancy were randomly divided into a control group, a model group, an EA group and an ibuprofen group, 10 rats in each group. PDM model was prepared by injection of estradiol benzoate and oxytocin. Except the control group, the rats in each group were subcutaneously injected with estradiol benzoate for 10 days, and oxytocin was injected on the 11th day. The rats in the EA group were intervened with EA (dense wave, frequency of 50 Hz) at "Guanyuan" (CV 4) and "Sanyinjiao" (SP 6) at the same time of modeling, once a day, 20 min each time, for 10 consecutive days. The rats in the ibuprofen group were treated with 0.8 mL of ibuprofen by gavage (concentration of ibuprofen solution was 1.25 mg/mL) for 10 consecutive days. After modeling, the writhing reaction was observed. After intervention, the HE staining method was used to observe the histological morphology of uterus and evaluate the pathological damage score of uterus; ELISA method was used to detect the serum levels of prostaglandin E2 (PGE2) and prostaglandin F2α (PGF2α); Western blot method was used to detect the protein expression of NLRP3, apoptosis related spot like protein (ASC), caspase-1, GSDMD, GSDMD-N and inflammatory factors (interleukin [IL]-1β, IL-18) in uterine tissue.@*RESULTS@#In the model group, a large number of vacuolar degeneration and death of endometrial epithelial cells, spiral arterioles congestion in lamina propria and neutrophil infiltration were observed. In the EA group, there was a small amount of vacuolar degeneration and death of endometrial epithelial cells, a small amount of spiral arterioles congestion in the lamina propria, and a small amount of neutrophils infiltration. In the ibuprofen group, there was very small number of degeneration and death of endometrial epithelial cells, and no obvious arterial congestion was found in lamina propria, and neutrophil infiltration was occasionally seen. Compared with the control group, in the model group the number of writhing was increased (P<0.01), the writhing reaction score and serum level of PGF2α and PGF2α/PGE2 value were increased (P<0.01), the level of PGE2 was decreased (P<0.01). Compared with the model group, in the EA group and the ibuprofen group the number of writhing were decreased (P<0.05), the latency of writhing was prolonged (P<0.01), the writhing reaction scores and serum levels of PGF2α and PGF2α/PGE2 values were decreased (P<0.05, P<0.01), the levels of PGE2 were increased (P<0.01). Compared with the control group, the protein expression of NLRP3, ASC, caspase-1, GSDMD, GSDMD-N, IL-1β and IL-18 in the uterine tissues of rats was increased in the model group (P<0.01). Compared with the model group, the protein expression of NLRP3, ASC, caspase-1, GSDMD, GSDMD-N, IL-1β and IL-18 in the uterine tissues of rats was decreased in the EA group and the ibuprofen group (P<0.01, P<0.05). There was no significant difference between the EA group and the ibuprofen group in the above indexes (P>0.05).@*CONCLUSION@#EA could alleviate pain and uterine tissue injury in rats with PDM. The mechanism may be related to the inhibition of the activation of NLRP3 inflammasome in rat uterine tissues, thereby inhibiting pyroptosis and its inflammatory factors release.
Subject(s)
Animals , Female , Pregnancy , Rats , Caspases , Dinoprost , Dinoprostone , Dysmenorrhea , Electroacupuncture , Ibuprofen , Inflammasomes , Interleukin-18 , NLR Family, Pyrin Domain-Containing 3 Protein , Oxytocin , Phosphate-Binding Proteins , Pyroptosis , Rats, Sprague-Dawley , Uterus摘要
Emerging SARS-CoV-2 variants have made COVID-19 convalescents susceptible to re-infection and have raised concern about the efficacy of inactivated vaccination in neutralization against emerging variants and antigen-specific B cell response. To this end, a study on a long-term cohort of 208 participants who have recovered from COVID-19 was conducted, and the participants were followed up at 3.3 (Visit 1), 9.2 (Visit 2), and 18.5 (Visit 3) months after SARS-CoV-2 infection. They were classified into three groups (no-vaccination (n = 54), one-dose (n = 62), and two-dose (n = 92) groups) on the basis of the administration of inactivated vaccination. The neutralizing antibody (NAb) titers against the wild-type virus continued to decrease in the no-vaccination group, but they rose significantly in the one-dose and two-dose groups, with the highest NAb titers being observed in the two-dose group at Visit 3. The NAb titers against the Delta variant for the no-vaccination, one-dose, and two-dose groups decreased by 3.3, 1.9, and 2.3 folds relative to the wild-type virus, respectively, and those against the Omicron variant decreased by 7.0, 4.0, and 3.8 folds, respectively. Similarly, the responses of SARS-CoV-2 RBD-specific B cells and memory B cells were boosted by the second vaccine dose. Results showed that the convalescents benefited from the administration of the inactivated vaccine (one or two doses), which enhanced neutralization against highly mutated SARS-CoV-2 variants and memory B cell responses. Two doses of inactivated vaccine among COVID-19 convalescents are therefore recommended for the prevention of the COVID-19 pandemic, and vaccination guidelines and policies need to be updated.
摘要
Emerging SARS-CoV-2 variants have made COVID-19 convalescents susceptible to re-infection and have raised concern about the efficacy of inactivated vaccination in neutralization against emerging variants and antigen-specific B cell response. To this end, a study on a long-term cohort of 208 participants who have recovered from COVID-19 was conducted, and the participants were followed up at 3.3 (Visit 1), 9.2 (Visit 2), and 18.5 (Visit 3) months after SARS-CoV-2 infection. They were classified into three groups (no-vaccination (n = 54), one-dose (n = 62), and two-dose (n = 92) groups) on the basis of the administration of inactivated vaccination. The neutralizing antibody (NAb) titers against the wild-type virus continued to decrease in the no-vaccination group, but they rose significantly in the one-dose and two-dose groups, with the highest NAb titers being observed in the two-dose group at Visit 3. The NAb titers against the Delta variant for the no-vaccination, one-dose, and two-dose groups decreased by 3.3, 1.9, and 2.3 folds relative to the wild-type virus, respectively, and those against the Omicron variant decreased by 7.0, 4.0, and 3.8 folds, respectively. Similarly, the responses of SARS-CoV-2 RBD-specific B cells and memory B cells were boosted by the second vaccine dose. Results showed that the convalescents benefited from the administration of the inactivated vaccine (one or two doses), which enhanced neutralization against highly mutated SARS-CoV-2 variants and memory B cell responses. Two doses of inactivated vaccine among COVID-19 convalescents are therefore recommended for the prevention of the COVID-19 pandemic, and vaccination guidelines and policies need to be updated.
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Nuclear transporter importin-β1 is emerging as an attractive target by virtue of its prevalence in many cancers. However, the lack of druggable inhibitors restricts its therapeutic proof of concept. In the present work, we optimized a natural importin-β1 inhibitor DD1 to afford an improved analog DD1-Br with better tolerability (>25 folds) and oral bioavailability. DD1-Br inhibited the survival of castration-resistant prostate cancer (CRPC) cells with sub-nanomolar potency and completely prevented tumor growth in resistant CRPC models both in monotherapy (0.5 mg/kg) and in enzalutamide-combination therapy. Mechanistic study revealed that by targeting importin-β1, DD1-Br markedly inhibited the nuclear accumulation of multiple CRPC drivers, particularly AR-V7, a main contributor to enzalutamide resistance, leading to the integral suppression of downstream oncogenic signaling. This study provides a promising lead for CRPC and demonstrates the potential of overcoming drug resistance in advanced CRPC via targeting importin-β1.
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The purpose of this study was to establish a suitable method for extracting cerebrospinal fluid (CSF) from C57BL/6 mice. A patch clamp electrode puller was used to draw a glass micropipette, and a brain stereotaxic device was used to fix the mouse's head at an angle of 135° from the body. Under a stereoscopic microscope, the skin and muscle tissue on the back of the mouse's head were separated, and the dura mater at the cerebellomedullary cistern was exposed. The glass micropipette (with an angle of 20° to 30° from the dura mater) was used to puncture at a point 1 mm inboard of Y-shaped dorsal vertebral artery for CSF sampling. After the first extraction, the glass micropipette was connected with a 1 mL sterile syringe to form a negative pressure device for the second extraction. The results showed that the successful rate of CSF extraction was 83.33% (30/36). Average CSF extraction amount was (7.16 ± 0.43) μL per mouse. In addition, C57BL/6 mice were given intranasally ferric ammonium citrate (FAC) to establish a model of brain iron accumulation, and the CSF extraction technique established in the present study was used for sampling. The results showed that iron content in the CSF from the normal saline control group was not detected, while the iron content in the CSF from FAC-treated group was (76.24 ± 38.53) μmol/L, and the difference was significant. These results suggest that glass micropipette vacuum technique of CSF sampling established in the present study has the advantages of simplicity, high success rate, large extraction volume, and low bleeding rate, and is suitable for the research on C57BL/6 mouse neurological disease models.
Subject(s)
Mice , Animals , Vacuum , Mice, Inbred C57BL , Cisterna Magna , Brain , Cerebrospinal Fluid摘要
It has been well documented that exercise can improve bone metabolism, promote bone growth and development, and alleviate bone loss. MicroRNAs (miRNAs) are widely involved in the proliferation and differentiation of bone marrow mesenchymal stem cells, osteoblasts, osteoclasts and other bone tissue cells, and regulation of balance between bone formation and bone resorption by targeting osteogenic factors or bone resorption factors. Thus miRNAs play an important role in the regulation of bone metabolism. Recently, regulation of miRNAs are shown to be one of the ways by which exercise or mechanical stress promotes the positive balance of bone metabolism. Exercise induces changes of miRNAs expression in bone tissue and regulates the expression of related osteogenic factors or bone resorption factors, to further strengthen the osteogenic effect of exercise. This review summarizes relevant studies on the mechanism whereby exercise regulates bone metabolism via miRNAs, providing a theoretical basis for osteoporosis prevention and treatment with exercise.
Subject(s)
Humans , MicroRNAs/metabolism , Osteogenesis/genetics , Cell Differentiation , Osteoblasts , Bone Resorption/metabolism摘要
ObjectiveTo explore the effect of tuina manipulation with different cervical rotation angle on carotid atherosclerosis. MethodsTwenty-five New Zealand rabbits were randomly selected， 5 of which as the control group， and the other 20 rabbits as the modeling group. The modeling group were made by arterial intimal balloon injury combined with high-fat diet， and were randomly divided into model group， cervical rotation angle of 90° group， cervical rotation angle of 105° group and cervical rotation angle of 120° group （5 rabbits in each group） after successful modeling. After relaxing the neck muscles of rabbits with manipulation of one-finger meditation， rolling and dialing， the cervical vertebrae of each group was rotated by 90°， 105°and 120° respectively for 2 weeks， while the other two groups were not intervened. Then took the entire length of the left common carotid artery from the rabbit， observed the pathological morphology of the carotid artery tissue using HE staining， and observed the expression of CD68 and MMP-2 in the carotid artery tissue using immunohistochemistry staining， and conducted semi quantitative analysis. ResultsHE staining showed that there was no obvious pathological change in the carotid artery in the control group； the model group showed subintimal foam cells gathered， vascular smooth muscle cells increased and arranged disorderly， and some vascular smooth muscle cells entered the intima from the media to form a fibrous cap. Compared with the model group， the number of foam cells increased and the symptoms of lumen stenosis were improved in all groups of cervical rotation； compared with the group with 90°rotation， the endothelium tended to fall off slightly in the group with 105°rotation and 120°rotation. Compared with the control group， the model group and the cervical rotation angle of 90° group ， cervical rotation angle of 105° group and cervical rotation angle of 120° group showed an increase in positive expression of CD68 and MMP-2 （P＜0.05）； compared with the model group， the positive expression of CD68 and MMP-2 in the cervical rotation angle of 90° group， cervical rotation angle of 105° group and cervical rotation angle of 120° group decreased （P＜0.05）； there was no statistically significant difference in CD68 positive expression among cervical rotation angle of 90° group， cervical rotation angle of 105° group and cervical rotation angle of 120° group compared in pairs （P＞0.05）； the positive expression of MMP-2 in the group of cervical rotation angle of 105° group and cervical rotation angle of 120 °group was higher than that in cervical rotation angle of 90° group （P＜0.05）. ConclusionTuina manipulation can promote the stability of plaque by reducing the CD68 content of macrophages and the level of MMP-2 in plaque； but as the rotation angle of the cervical spine increased， the expression of MMP-2 may increase， which may reduce the plaque stability to a certain extent and affect the therapeutic effect of tuina.
摘要
Radiation enteritis (RE) is a common syndrome in the radiotherapy of abdominal and pelvic malignant tumors, heavy influencing living quality, but no specific clinical regimens are available. Long oil (LO) is composed of the fat components from cuttlebone, safflower, walnut oil and rapeseed oil and has been clinically used for wound healing. In this study, oral LO was applied for the prevention and treatment of RE and the mechanisms were explored. Animal experiments were approved by the Ethics Committee of the Beijing Institute of Radiation Medicine, Academy of Military Medical Sciences, and the experiments were conducted in accordance with relevant guidelines and regulations. An RE mouse model was established after single whole abdominal γ-ray radiation of 13 Gy. LO (8 mL·kg-1) was intragastrically administered to the mice 1 h pre-radiation. Compared to the models, the mice of the LO group had more regenerated intestinal crypts and longer villus on day 3.5, and remarkable increase in the abundance of gut microbiota on day 7, especially the amounts of probiotics including Eubacterium and Lactobacillus. Moreover, the mice of the LO group showed longer total movement distance, shorter immobility time, and higher speed than the model mice on day 7. On day 14, the mice of the LO group showed the high descending of proinflammatory factors including tumor necrosis factor-α and interleukin-6, close to the normal levels. Therefore, oral LO can alleviate the inflamed syndromes of RE and improve the repair of damaged intestinal tissues. Moreover, the mice of the LO group had highly low permeability of intestinal mucosa according to the fluorescence labeling experiment, which was close to the normal level. Oral LO can protect intestine mucosa and prevent RE by modification of the intestinal microenvironment, alleviation of the inflammatory response, and promotion of tissue repair.
摘要
Objective: To prepare graphene oxide (GO)-containing gelatin methacrylate anhydride (GelMA) hydrogel and to investigate the effects of in situ photopolymerized GO-GelMA composite hydrogel in wound vascularization of full-thickness skin defect in mice. Methods: The experimental study method was used. The 50 μL of 0.2 mg/mL GO solution was evenly applied onto the conductive gel, and the structure and size of GO were observed under field emission scanning electron microscope after drying. Human skin fibroblasts (HSFs) were divided into 0 μg/mL GO (without GO solution, the same as below) group, 0.1 μg/mL GO group, 1.0 μg/mL GO group, 5.0 μg/mL GO group, and 10.0 μg/mL GO group treated with GO of the corresponding final mass concentration, and the absorbance value was detected using a microplate analyzer after 48 h of culture to reflect the proliferation activity of cells (n=6). HSFs and human umbilical vein vascular endothelial cells (HUVECs) were divided into 0 μg/mL GO group, 0.1 μg/mL GO group, 1.0 μg/mL GO group, and 5.0 μg/mL GO group treated with GO of the corresponding final mass concentration, and the migration rates of HSFs at 24 and 36 h after scratching (n=5) and HUVECs at 12 h after scratching (n=3) were detected by scratch test, and the level of vascular endothelial growth factor (VEGF) secreted by HSFs after 4, 6, and 8 h of culture was detected by enzyme-linked immunosorbent assay method (n=3). The prepared GO-GelMA composite hydrogels containing GO of the corresponding final mass concentration were set as 0 μg/mL GO composite hydrogel group, 0.1 μg/mL GO composite hydrogel group, 1.0 μg/mL GO composite hydrogel group, and 5.0 μg/mL GO composite hydrogel group to observe their properties before and after cross-linking, and to detect the release of GO after soaking with phosphate buffer solution for 3 and 7 d (n=3). The full-thickness skin defect wounds were made on the back of 16 6-week-old female C57BL/6 mice. The mice treated with in situ cross-linked GO-GelMA composite hydrogel containing GO of the corresponding final mass concentration were divided into 0 μg/mL GO composite hydrogel group, 0.1 μg/mL GO composite hydrogel group, 1.0 μg/mL GO composite hydrogel group, and 5.0 μg/mL GO composite hydrogel group according to the random number table, with 4 mice in each group. The general condition of wound was observed and the wound healing rate was calculated on 3, 7, and 14 d of treatment, the wound blood perfusion was detected by laser Doppler flowmetry on 3, 7, and 14 d of treatment and the mean perfusion unit (MPU) ratio was calculated, and the wound vascularization on 7 d of treatment was observed after hematoxylin-eosin staining and the vascular density was calculated (n=3). The wound tissue of mice in 0 μg/mL GO composite hydrogel group and 0.1 μg/mL GO composite hydrogel group on 7 d of treatment was collected to observe the relationship between the distribution of GO and neovascularization by hematoxylin-eosin staining (n=3) and the expression of VEGF by immunohistochemical staining. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and Tukey's method. Results: GO had a multilayered lamellar structure with the width of about 20 μm and the length of about 50 μm. The absorbance value of HSFs in 10.0 μg/mL GO group was significantly lower than that in 0 μg/mL GO group after 48 h of culture (q=7.64, P<0.01). At 24 h after scratching, the migration rates of HSFs were similar in the four groups (P>0.05); at 36 h after scratching, the migration rate of HSFs in 0.1 μg/mL GO group was significantly higher than that in 0 μg/mL GO group, 1.0 μg/mL GO group, and 5.0 μg/mL GO group (with q values of 7.48, 10.81, and 10.20, respectively, P<0.01). At 12 h after scratching, the migration rate of HUVECs in 0.1 μg/mL GO group was significantly higher than that in 0 μg/mL GO group, 1.0 μg/mL GO group, and 5.0 μg/mL GO group (with q values of 7.11, 8.99, and 14.92, respectively, P<0.01), and the migration rate of HUVECs in 5.0 μg/mL GO group was significantly lower than that in 0 μg/mL GO group and 1.0 μg/mL GO group (with q values of 7.81 and 5.33, respectively, P<0.05 or P<0.01 ). At 4 and 6 h of culture, the VEGF expressions of HSFs in the four groups were similar (P>0.05); at 8 h of culture, the VEGF expression of HSFs in 0.1 μg/mL GO group was significantly higher than that in 0 μg/mL GO group and 5.0 μg/mL GO group (with q values of 4.75 and 4.48, respectively, P<0.05). The GO-GelMA composite hydrogels in the four groups were all red liquid before cross-linking, which turned to light yellow gel after cross-linking, with no significant difference in fluidity. The GO in the GO-GelMA composite hydrogel of 0 μg/mL GO composite hydrogel group had no release of GO at all time points; the GO in the GO-GelMA composite hydrogels of the other 3 groups was partially released on 3 d of soaking, and all the GO was released on 7 d of soaking. From 3 to 14 d of treatment, the wounds of mice in the 4 groups were covered with hydrogel dressings, kept moist, and gradually healed. On 3, 7, and 14 d of treatment, the wound healing rates of mice in the four groups were similar (P>0.05). On 3 d of treatment, the MPU ratio of wound of mice in 0.1 μg/mL GO composite hydrogel group was significantly higher than that in 0 μg/mL GO composite hydrogel group, 1.0 μg/mL GO composite hydrogel group, and 5.0 μg/mL GO composite hydrogel group (with q values of 10.70, 11.83, and 10.65, respectively, P<0.05 or P<0.01). On 7 and 14 d of treatment, the MPU ratios of wound of mice in the four groups were similar (P>0.05). The MPU ratio of wound of mice in 0.1 μg/mL GO composite hydrogel group on 7 d of treatment was significantly lower than that on 3 d of treatment (q=14.38, P<0.05), and that on 14 d of treatment was significantly lower than that on 7 d of treatment (q=27.78, P<0.01). On 7 d of treatment, the neovascular density of wound of mice on 7 d of treatment was 120.7±4.1 per 200 times of visual field, which was significantly higher than 61.7±1.3, 77.7±10.2, and 99.0±7.9 per 200 times of visual field in 0 μg/mL GO composite hydrogel group, 1.0 μg/mL GO composite hydrogel group, and 5.0 μg/mL GO composite hydrogel group (with q values of 12.88, 7.79, and 6.70, respectively, P<0.01), and the neovascular density of wound of mice in 1.0 μg/mL GO composite hydrogel group and 5.0 μg/mL GO composite hydrogel group was significantly higher than that in 0 μg/mL GO composite hydrogel group (with q values of 5.10 and 6.19, respectively, P<0.05). On 7 d of treatment, cluster of new blood vessels in wound of mice in 0.1 μg/mL GO composite hydrogel group was significantly more than that in 0 μg/mL GO composite hydrogel group, and the new blood vessels were clustered near the GO; a large amount of VEGF was expressed in wound of mice in 0.1 μg/mL GO composite hydrogel group in the distribution area of GO and new blood vessels. Conclusions: GO with mass concentration lower than 10.0 μg/mL had no adverse effect on proliferation activity of HSFs, and GO of 0.1 μg/mL can promote the migration of HSFs and HUVECs, and can promote the secretion of VEGF in HSFs. In situ photopolymerized of GO-GelMA composite hydrogel dressing can promote the wound neovascularization of full-thickness skin defect in mice and increase wound blood perfusion in the early stage, with GO showing an enrichment effect on angiogenesis, and the mechanism may be related to the role of GO in promoting the secretion of VEGF by wound cells.
Subject(s)
Animals , Female , Humans , Mice , Anhydrides , Endothelial Cells , Eosine Yellowish-(YS) , Gelatin/pharmacology , Graphite , Hematoxylin , Hydrogels/pharmacology , Methacrylates , Mice, Inbred C57BL , Neovascularization, Pathologic , Skin Abnormalities , Vascular Endothelial Growth Factor A摘要
Objective To study the correlation between regional cerebral glucose metabolism and behavioral scores in middle cerebral artery occlusion(MCAO)model rats before and after the intervention of constraint induced movement therapy(CIMT),and the correlation between the natural recovery processand motor function recovery in MCAO model rats and the brain activation after CIMT intervention,and to further explore the mechanism of CIMT. Methods Twenty-two adult male Sprague-Dawley(SD)rats were randomly divided into an ischemic group treated with CIMT (CIMT,n=6),an ischemic group (Control,n=6),a sham-operated group(Sham,n=6),and a blank control group(Normal,n=4). The MCAO models of rats in the CIMT group and Control group were established by thread embolism method. The middle cerebral artery was not blocked during the operation for the Sham group,and the Normal group was not given any special treatment. After operation,rats in the CIMT group and Sham group were treated with CIMT. On the 7th day(d7)and the 22nd day(d22)after surgery,foot-fault test(FFT)and the beam balance and walking (BBW) test were used to evaluate the forelimb motor;micro positron emission tomography-computed tomography (micro PET/CT) imaging with fluorodeoxyglucose (18F-FDG) was used to scan the glucose metabolism in different brain regions of rats;Pearson correlation analysis was used to analyze the correlation between behavioral scores and glucose metabolism level in the CIMT group and Control group. Results On d7,the BBW score in the CIMT group and Control group was negatively correlated with glucose metabolism in the left insular cortex and the auditory cortex,and positively correlated with glucose metabolism in the right posterior hippocampus,superior colliculus,and inferior colliculus,with statistically significant differences;the FFT score was negatively correlated with glucose metabolism in the left somatosensory cortex, insular cortex and orbitofrontal cortex, and positively correlated with glucose metabolism in the right midbrain,with statistically significant differences. On d22, the BBW score in the CIMT group and Control group was positively correlated with glucose metabolism in the amygdala,caudate putamen,insular cortex and entorhinal cortex,and negatively correlated with glucose metabolism in the nucleus accumbens (Acb) core shell and caudate putamen in the right brain region,with statistically significant differences;the FFT score was negatively correlated with the entorhinal cortex in the right hemisphere and the difference was statistically significant. Conclusion The recovery of motor function promoted by CIMT was associated with the activation of both cerebral hemispheres in rats. The improvement of balance function promoted by CIMT in rats with cerebral ischemia was mainly related to the activation of Acb core shell in the right hemisphere. The recovery of fine grasping function promoted by CIMT may be related to the activation of the right entorhinal cortex.