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The evaluation of germplasm resources is the prerequisite for the development, utilization, and conservation of Chinese medicinal resources. The selection of excellent germplasm is the key to the breeding and orderly production of Pinellia ternata. In this study, 21 germplasm materials of P. ternata from major production areas in China were collected and analyzed for population diversity after phenotypic preliminary screening. The results have revealed that the P. ternata population has abundant phenotypic variation, and the phenotypic changes could be divided into five phenotypes in terms of organ trait variation. Further analysis of variation in 20 quantitative traits of the population revealed that the coefficient of variation for adenosine content(339.05%) was the largest, while the coefficient of variation for the underground plant height(16.35%) was the smallest. Correlation analysis showed that there was a strong correlation among various traits, with 52 pairs of traits showing highly significant correlation(P<0.01) and 19 pairs of traits showing a significant correlation(P<0.05). The 21 germplasms in the test could be classified into three major clusters by cluster analysis, with Cluster Ⅱ having the highest number and content of nucleosides, making it suitable for the selection and breeding of P. ternata varieties with high content of nucleosides. The yield in Cluster Ⅲ was higher than that in other groups, making it suitable for the selection and breeding of P. ternata varieties with a high yield. All trait indicators could be simplified into five principal component factors through principal component analysis, and the cumulative contribution rate was up to 86.04%. Further, comprehensive analysis using membership function and stepwise regression analysis identified nine traits, such as plant height, main leaf length, and underground plant height as characteristic indicators for the comprehensive evaluation of germplasm resources of P. ternata. BX007, BX008, and BX005 were identified as germplasms with both high yield and high uridine content, with BX007 having the highest uridine content of 479.51 μg·g~(-1). It belonged to the germplasm of P. ternata with double bulbils and could be cultivated as a potential good variety. Based on the phenotypic classification of P. ternata, systematic resource evaluation was carried out in this study, which could lay a foundation for the excavation of genetic resources and the breeding of new varieties of P. ternata.
Subject(s)
Plants, Medicinal , Pinellia/genetics , Plant Breeding , Phenotype , UridineABSTRACT
Background and Objectives@#Endothelial progenitor cells (EPCs) and endothelial cells (ECs) have been applied in the clinic to treat pulmonary arterial hypertension (PAH), a disease characterized by disordered pulmonary vasculature. However, the lack of sufficient transplantable cells before the deterioration of disease condition is a current limitation to apply cell therapy in patients. It is necessary to differentiate pluripotent stem cells (PSCs) into EPCs and identify their characteristics. @*Methods@#and Results: Comparing previously reported methods of human PSCs-derived ECs, we optimized a highly efficient differentiation protocol to obtain cells that match the phenotype of isolated EPCs from healthy donors. The protocol is compatible with chemically defined medium (CDM), it could produce a large number of clinically applicable cells with low cost. Moreover, we also found PSCs-derived EPCs express CD133, have some characteristics of mesenchymal stem cells and are capable of homing to repair blood vessels in zebrafish xenograft assays. In addition, we further revealed that IPAH PSCs-derived EPCs have higher expression of proliferation-related genes and lower expression of immune-related genes than normal EPCs and PSCs-derived EPCs through microarray analysis. @*Conclusions@#In conclusion, we optimized a highly efficient differentiation protocol to obtain PSCs-derived EPCs with the phenotypic and molecular characteristics of EPCs from healthy donors which distinguished them from EPCs from PAH.
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ObjectiveTo explore the mechanism of antidepressant effect of lily polysaccharide （LLP）and astragalus polysaccharide（APS）. MethodSixty KM mice were randomly divided into blank group， model group， fluoxetine hydrochloride （8 mg·kg-1）group， LLP （0.2 g·kg-1）group， APS （0.2 g·kg-1）group and polysaccharide combination （LLP+APS，0.1 g·kg-1+0.1 g·kg-1）group， with 10 mice in each group. Except the blank group， the other groups were given chronic unpredictable mild stress （CUMS） induced mouse depression model. On the 29th day of modeling，fluoxetine hydrochloride group was given corresponding dose of fluoxetine hydrochloride， and polysaccharide groups were given corresponding drug. The depressive behavior of mice was evaluated by behavioral indexes such as body mass change， open field test. The morphological changes of hippocampal CA1 neurons were observed by Nissl staining. The contents of 5-hydroxytryptamine （5-HT）， adrenocorticotropic hormone （ACTH）， and corticosterone （CORT）， in brain tissue and plasma were measured by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression levels of related proteins in adenylate cyclase/cyclic adenylate phosphate/protein kinase A （AC/cAMP/PKA） signal pathway. ResultCompared with the blank group， mice in the model group gained weight slowly， the total distance， central distance and sugar water preference rate decreased significantly （P<0.01）， the depressive behavior was significant， the hippocampal neurons were seriously damaged， the content of 5-HT decreased （P<0.01）， the contents of ACTH and CORT increased significantly （P<0.01）， adenylate cyclase 6（ADCY6）， PKA and cAMP response element binding protein-1 （CREB-1） and brain-derived neurotrophic factor （BDNF） protein expression decreased significantly （P<0.01）. Compared with the model group， depressive behavior of mice in LLP group， APS group and LLP+APS group was significantly improved （P<0.01）. The antidepressant effect of LLP+APS was better than that of LLP and APS. Each administration group could alleviate the damage of hippocampal neurons in varying degrees， significantly increase the content of 5-HT in brain tissue （P<0.01）， and reduce the levels of ACTH and CORT in plasma （P<0.05）. The protein levels of ADCY6， PKA， CREB-1 and BDNF were significantly increased （P<0.05）. ConclusionThe antidepressant effect of LLP+APS is significantly enhanced and has a synergistic effect. The mechanism may be closely related to affecting the content of neurotransmitters， inhibiting HPA axis activity and activating AC/cAMP/PKA signal transduction pathway.
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Objective:To determine the proportion of tobacco smoking and tobacco dependence among residents in Changning District of Shanghai， and provide scientific evidence for future intervention. Methods:A questionnaire survey was conducted among 1 760 residents over 15 years old in Changning District， covering smoking behavior， tobacco dependence and smoking relapse. Chi-square test was used to compare the proportions of smoking-related behavior among different groups， and logistic regression model was used to determine the risk factors. Results:Smoking proportion among residents over 15 years old in Changning District was determined to be 17.0%（299/1 760）， which was 35.8% （285/795） for males and 1.5% （14/965） for females. The smoking proportion was significantly higher in the residents being male， older， with less education and underlying diseases （P<0.05）. The residents "knowing the hazard of smoking but remaining smoking" was the most serious tobacco dependence， which was determined to be 85.3% （255/299） among the smokers. The most common symptom leading to smoking relapse was "impulse to smoking " （61.1%，138/226）. Among them， "smoking impulse"（χ2=8.512，P=0.004）， " restlessness "（χ2=4.000，P=0.045）， and "depression" （χ2=6.328，P=0.010） are the three symptoms which had an effect on whether the smokers relapsed. Conclusion:The smoking rate in Changning District of Shanghai is lower than the national and local average. Tobacco dependence mainly focuses on awareness and attitude. Intervention should focus on the key population with high smoking rate. We should correct the severe tobacco dependence， intervene the main adverse symptoms， and guide smoking cessation. In addition， we should provide more accurate and effectives smoking cessation services based on health communication.
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BACKGROUND@#The significant morbidity and mortality resulted from the infection of a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) call for urgent development of effective and safe vaccines. We report the immunogenicity and safety of an inactivated SARS-CoV-2 vaccine, KCONVAC, in healthy adults.@*METHODS@#Phase 1 and phase 2 randomized, double-blind, and placebo-controlled trials of KCONVAC were conducted in healthy Chinese adults aged 18 to 59 years. The participants in the phase 1 trial were randomized to receive two doses, one each on Days 0 and 14, of either KCONVAC (5 or 10 μg/dose) or placebo. The participants in the phase 2 trial were randomized to receive either KCONVAC (at 5 or 10 μg/dose) or placebo on Days 0 and 14 (0/14 regimen) or Days 0 and 28 (0/28 regimen). In the phase 1 trial, the primary safety endpoint was the proportion of participants experiencing adverse reactions/events within 28 days following the administration of each dose. In the phase 2 trial, the primary immunogenicity endpoints were neutralization antibody seroconversion and titer and anti-receptor-binding domain immunoglobulin G seroconversion at 28 days after the second dose.@*RESULTS@#In the phase 1 trial, 60 participants were enrolled and received at least one dose of 5-μg vaccine (n = 24), 10-μg vaccine (n = 24), or placebo (n = 12). In the phase 2 trial, 500 participants were enrolled and received at least one dose of 5-μg vaccine (n = 100 for 0/14 or 0/28 regimens), 10-μg vaccine (n = 100 for each regimen), or placebo (n = 50 for each regimen). In the phase 1 trial, 13 (54%), 11 (46%), and seven (7/12) participants reported at least one adverse event (AE) after receiving 5-, 10-μg vaccine, or placebo, respectively. In the phase 2 trial, 16 (16%), 19 (19%), and nine (18%) 0/14-regimen participants reported at least one AE after receiving 5-, 10-μg vaccine, or placebo, respectively. Similar AE incidences were observed in the three 0/28-regimen treatment groups. No AEs with an intensity of grade 3+ were reported, expect for one vaccine-unrelated serious AE (foot fracture) reported in the phase 1 trial. KCONVAC induced significant antibody responses; 0/28 regimen showed a higher immune responses than that did 0/14 regimen after receiving two vaccine doses.@*CONCLUSIONS@#Both doses of KCONVAC are well tolerated and able to induce robust immune responses in healthy adults. These results support testing 5-μg vaccine in the 0/28 regimen in an upcoming phase 3 efficacy trial.@*TRIAL REGISTRATION@#http://www.chictr.org.cn/index.aspx (No. ChiCTR2000038804, http://www.chictr.org.cn/showproj.aspx?proj=62350; No. ChiCTR2000039462, http://www.chictr.org.cn/showproj.aspx?proj=63353).
Subject(s)
Adult , Humans , COVID-19 , COVID-19 Vaccines , Double-Blind Method , SARS-CoV-2 , Vaccines, Inactivated/adverse effectsABSTRACT
OBJECTIVE@#To observe the effect of electroacupuncture on postpartum uterine contraction pain and uterine involution.@*METHODS@#A total of 80 patients with postpartum uterine contraction pain were randomly divided into an observation group and a control group, 40 cases in each group. The observation group was treated with electroacupuncture at Dahe (KI 12), Zhongzhu (KI 15), Hegu (LI 4), Xuehai (SP 10), etc. for 30 min, once a day, 3 days were as one course, and 2 courses with 1-day interval were required. The control group was treated with oral @*RESULTS@#Compared before treatment, the VAS scores of 24, 48, 72 h into treatment and after treatment were decreased in both groups (@*CONCLUSION@#Electroacupuncture can effectively relieve postpartum uterine contraction pain, accelerate the discharge of residual uterine hemorrhage in the uterine cavity, and promote uterine involution.
Subject(s)
Female , Humans , Pregnancy , Acupuncture Points , Electroacupuncture , Pain , Postpartum Period , Uterine ContractionABSTRACT
OBJECTIVE@#To compare the clinical therapeutic effect on painful diabetic peripheral neuropathy (PDPN) between dragon-tiger fighting needling and pregabalin capsules.@*METHODS@#A total of 60 patients with PDPN were randomized into an observation group and a control group, 30 cases in each one. On the base of treatment with routine anti-hyperglycaemic measures and nutritional neurotherapy, the dragon-tiger fighting needling was exerted at Sanyinjiao (SP 6), Zusanli (ST 36), Yinlingquan (SP 9) and Xuehai (SP 10) in the observation group, once daily. Pregabalin capsules were prescribed for oral administration in the control group, 75 mg, twice a day. The treatment for 2 weeks was as one course and 2 courses of treatment were required in total. The score of visual analogue scale (VAS), the score of MOS item short form health survey (SF-36) and nerve conduction velocity before and after treatment were compared between the two groups. The clinical therapeutic effect was evaluated in the two groups.@*RESULTS@#After treatment, VAS score was reduced as compared with before treatment in the two groups (@*CONCLUSION@#The dragon-tiger fighting needling therapy relieves painful symptoms, improves the quality of life and increases nerve conduction velocity in the patients with diabetic peripheral neuropathy, and the therapeutic effect is better than oral administration of pregabalin capsules.
Subject(s)
Animals , Humans , Acupuncture Points , Acupuncture Therapy , Diabetes Mellitus , Diabetic Neuropathies/drug therapy , Quality of Life , Tigers , Treatment OutcomeABSTRACT
To investigate the potential toxic effects and mechanisms of Tris(1; 3-dichloro-2-propyl) phosphate (TDCIPP) on thyroid in female SD rats. Thirty-two 3-weeks-old female SD rats were randomly divided into normal group(treated with corn oil ), and low/moderate/high-dose group treated with TDCIPP (dissolved in corn oil )(n=8). All rats were treated with corn oil or TDCIPP (50, 100, 250 mg/(kg·d)) once a day during a 21-day period. All rats were sacrificed after the last administration. Serum thyroid stimulating hormone (TSH), 3,3',5-triiodothyronine (T3), 3,3',5,5'-tetraiodothyronine (T4), free 3,3',5,5'-tetraiodothyronine (FT4) were detected with ELISA kit. Morphology of thyroid was observed with hematoxylin and eosin (HE) staining. Expressions of genes and proteins correlate with thyroid were measured respectively by real-time fluorescence quantitative PCR and Western blot. Compared with control group, morphology of thyroid showed follicles irregular arrangement, hypocolloid, and follicular hyperplasia in TDCIPP treatment groups. The levels of serum TSH in low-dose TDCIPP group and T3 in high-dose TDCIPP group were significantly higher than those in control group(P＜0.05). Thyroid stimulating hormone receptor (TSHR) mRNA expression was decreased distinctly in low-dose TDCIPP group, while the expression of thyroperoxidase (TPO) mRNA was increased notably in moderate and high-dose TDCIPP groups(P＜0.05,P＜0.01). Compared with control group, the level of TRβ protein was decreased significantly in moderate and high-dose TDCIPP groups, while the expressions of udp-glucuronosyl-transferases (UGTs) and cytochrome-p450-3A1 (CYP3A1) proteins were upregulated notably in TDCIPP treatment groups(P＜0.05). Treated with 50 mg/(kg·d) TDCIPP can cause thyroid hyperplasia, change the levels of thyroid hormones, and disturb thyroid function, therefore, it has toxic effects on the thyroid.
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OBJECTIVE@#To observe the clinical effect of warm acupuncture on endometrial cavity fluid (ECF) from in vitro fertilization-embryo transfer (IVF-ET), and to explore the mechanism of warm acupuncture on ECF.@*METHODS@#Twenty-nine patients who were prepared for IVF-ET from 2016 to 2019 and whose transplantation was cancelled due to ECF found by vaginal B-ultrasound examination were divided into an observation group (14 cases) and a control group (15 cases) according to random number table method. The warm acupuncture was applied at Zhongwan (CV 12), Qihai (CV 6), Guanyuan (CV 4), Zhongji (CV 3), Guilai (ST 29), Zigong (EX-CA 1), Zusanli (ST 36), Sanyinjiao (SP 6) after the end of menstruation in the observation group, the treatment lasted for 60 min, once a day, 5 times as a course, with 2 days interval between the courses and 3 consecutive courses of treatment were given, until the embryo transfer was performed in the IVF assisted pregnancy cycle. After the end of menstruation, fresh leonurus japonicus capsule was given in the control group, 3 times a day, 0.8 g each time, 7 days as a course, and 3 courses of continuous treatment were received, until the embryo transfer was performed in the IVF assisted pregnancy cycle. The changes of ECF before and after treatment, the time required to prepare for embryo transfer during IVF assisted pregnancy cycle, and the clinical outcome of embryo transfer were observed in the two groups.@*RESULTS@#The decrease of ECF in the observation group was more significant than that in the control group (<0.05). The time required for the embryo transfer in the IVF assisted pregnancy cycle in the observation group was shorter than that in the control group (<0.05). The clinical pregnancy rate in the observation group was 42.9% (6/14), which was significantly higher than 26.7% (4/15) in the control group (<0.05).@*CONCLUSION@#Warm acupuncture may improve the clinical pregnancy rate by raising the local temperature of the lower abdomen, accelerating the blood circulation around the uterus and appendages, promoting the absorption of ECF, improving the uterine environment and endometrial receptivity.
Subject(s)
Female , Humans , Pregnancy , Acupuncture Therapy , Embryo Transfer , Endometrium , Fertilization in Vitro , Pregnancy RateABSTRACT
A sensing system based on AuNP-AuNP-UCNP triple structure for efficient detection of dual targets was constructed. In the preparation of triple structure, the gold nanoparticles (AuNPs) and upconversion nanoparticles (NaYF4: Yb, Er, Gd, UCNPs) were synthesized and surface modified. Then the two nanoparticles and their aptamers were connected to form two kinds of optical fluorescent probes. A nucleic acid sequence that matches with two aptamers was designed, rendering the probes to get close based on the principle of complementary base pairing. On the basis of this, a sensing system with a triple structure was prepared,and its connecting effect was characterized by TEM. With this system, dual targets of bisphenol A and estradiol were efficiently and conveniently detected through quantitative determination by fluorescence and UV spectrophotometer. At reaction temperature of 30℃ and pH=7.8,this method exhibited good linear range for determination of bisphenol A and estradiol from 2 ng/mL to 200 ng/mL and from 10 ng/mL to 150 ng/mL, with limits of detection of 0.2 ng/mL and 0.5 ng/mL, respectively. This sensing system with the triple structure owned better specificity to structural and functional analogues, and showed good repeatability and stability. What's more,this sensing system was applied in actual water detection,with the recoveries between 86.1% and 107. 4%, and the relative standard deviation below 6. 8%. This method showed promising applications in other environmental estrogens in water samples.
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The changes of endogenous metabolites in urine samples that come from pneumonia patients of 6 months to 6 years old children were analyzed by metabolomics methods based on gas chromatography and mass spectrometry (GC-MS).The aim of this study was to analyze and study the pathogenesis of endogenous metabolites in children with pneumonia and the pathogenesis of pneumonia susceptibility.The urine samples were collected and divided into normal children group (NC group,n=29),first infection with pneumonia group (FIP group,n=35),and repeated infection with pneumonia group (RIP group,n=31).The urine metabolic profile of pneumonia was obtained by GC-MS.Principal component analysis(PCA) and partial least squares-discriminant analysis (PLS-DA) were used to analyze the data.The results were analyzed by one-way analysis of variance and Fold change.Finally,there was significant difference between the normal group and the pneumonia group,the significant metabolites were serine,histidine,proline,norleucine,glutamine,stearic acid,valine,isoleucine with p value<0.05 and Fold change>5,and indole-3-acetic acid,creatine,ethanolamine,mannosylglycerol and fructose were significant between the two pneumonia groups with p value<0.05.The urinary metabolites demonstrated that amino acid metabolism and glucose metabolism were the main metabolic pathways and responsible for the susceptibility to pneumonia.
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A method for the determination of 15 kinds of nitroaromatics in aqueous samples was developed by dispersive liquid-liquid microextraction and gas chromatography with electron capture detection. A high-density extractant applied in electron capture detector was screened out. The chromatographic conditions, types and dosages of extractants, types and dosages of dispersants, extraction time and the extraction temperature were optimized. The results showed that DB-35 capillary column had the best separation performance for the 15 kinds of nitroaromatics. The nitroaromatics could be separated within 22 min using programmed temperature control as follows: holding at an initial temperature of 80℃ and then heating to 180℃ at a ramping rate of 5℃ / min. For the extraction of 15 kinds of nitroaromatics from 5 mL of aqueous sample, the extraction equilibrium could be reached within 30 s with a high extraction recovery of over 90%when using 100 μL of chlorobenzene as extracting solvent and 400 μL of methanol as disperser solvent. In addition, the enrichment factor could approach a high value of 45. 0 - 48. 8. The sediment collected by centrifugation was injected and analyzed by gas chromatography with electron capture detector. The limits of quantification of the developed method were 0. 03 - 0. 15 μg / L ( S / N = 10). The linear range was from 0. 20 μg / L to 50. 0 μg / L, while the correlation coefficients (R2 ) were more than 0. 998. At the spiked level of 0. 200 μg / L, the relative standard deviations of this method were 3. 3% -8. 9% , the relative recoveries ranged from 86. 0% to 103. 5% . At higher spiked level, the relative standard deviations were less than 5% , and the relative recoveries ranged from 94. 5% to 101. 5% .
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Objective To study the significance of the related parameters of the abduction angle ,anteversion angle and summit arc angle in the intraoperative localization of anterior column and anterior wall acetabular fractures .Methods The safety angle of abduction angle ,an-teversion angle,summit arc angle and anterior wall acetabular of 318 adult pelvic specimen(109 males and 109 females) were measured.The data was statistical analyzed.Results For the male pelvic specimen,the measured values of abduction angle of acetabulum was (43.5 ± 2.9)°, anteversion angle was (13.7 ±5.2)°,summit arc angle inside was(25.5 ±7.3)°,summit arc angle front was(37.8 ±7.2)°,sum-mit arc angle back(49.9 ±12.7)°.For female pelvic specimen,the measured values of abduction angles of acetabulum was (44.1 ±3.2)° and anteversion angle was (14.3 ±4.9)°,summit arc angle inside was(27.1 ±6.5)°,summit arc angle front was(36.3 ±9.2)°,summit arc angle back was(52.3 ±13.6)°.According to '2-7' sector method, the thickness of different point of anterior wall was measured .There were statistically significant differences in abduction angles ,anteversion angle and summit arc angle (P<0.05).And there was no statistically sig-nificant difference in the thickness of anterior column and anterior wall between the above three one angles (P>0.05),there was no signifi-cant difference in the acetabular abduction angle ,anteversion angle and summit arc angle between males and females .Conclusion The intra-operative localization of acetabulum abducted angle ,anteversion angle and summit arc angle in the acetabulum anterior column and anterior wall fractures can achieve satisfactory therapeutic effect .
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OBJECTIVE@#To observe the expression of phosphatase and tensin homology deleted on chromosome ten (PTEN) in myocardial tissue in patients with coronary heart disease, and explore the relevance between the expression of PTEN and the occurrence and development of coronary heart disease.@*METHODS@#A total of 16 death cases with pathological diagnosis of coronary heart disease were collected as experimental group, and 19 cases without myocardial lesions were selected as control group. The expression of PTEN protein and its mRNA were detected by immunohistochemistry and real-time fluorescence quantitative PCR respectively. The correlation between the expression of PTEN and the pathogenesis of coronary heart disease was analyzed.@*RESULTS@#The expression of PTEN protein in myocardium in cases with coronary heart disease was significantly lower compared with the control group (P < 0.05). There was no statistical difference of the expression of PTEN mRNA between experimental and control group (P > 0.05).@*CONCLUSION@#PTEN may be involved in the occurrence and development of coronary heart disease.
Subject(s)
Humans , Coronary Artery Disease/pathology , Myocardium/metabolism , PTEN Phosphohydrolase/metabolism , RNA, Messenger/metabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of recombinant adenovirus (phosphatidylinositol-3-kinases(PI3K)(I()-RNAi-AD which blocks the class I( PI3K signaling pathway on gastric carcinoma cells xenografts in nude mice.</p><p><b>METHODS</b>Subcutaneous tumor models of nude mice were established with SGC7901 cells and randomly divided into PI3K(I()-RNAi-AD group, NC-RNAi-GFP-AD group and control group. The tumor size and the inhibitory rate of tumor growth on days 3, 6, and 9 after cell transplantation were measured. The expression of TNF-α, COX2, P53, PCNA, E-cadherin and nm23/DNPK in tumor tissues were detected by immunohistochemistry.</p><p><b>RESULTS</b>Tumor growth was significantly inhibited in the PI3K(I()-RNAi-AD group(14.2%, 21.0%, and 28.1%) on days 3, 6, 9 compared with NC-RNAi-GFP-AD group(1.3%, 1.9%, and 2.0%, all P<0.05). The expressions of TNF-α, P53, E-cadherin and nm23/DNPK were up-regulated, and the expressions of COX2 and PCNA were down-regulated in the PI3K(I()-RNAi-AD group by immunohistochemical staining(all P<0.05).</p><p><b>CONCLUSIONS</b>PI3K(I()-RNAi-AD can inhibit the growth of SGC7901 cell transplantation tumor in vivo in nude mice by inhibiting cell growth, reducing the capacity of tumor invasion and inhibiting tumor angiogenesis.</p>
Subject(s)
Animals , Humans , Mice , Adenoviridae , Cell Line, Tumor , Cell Proliferation , Class I Phosphatidylinositol 3-Kinases , Heterografts , Mice, Inbred BALB C , Mice, Nude , Phosphatidylinositol 3-Kinases , Phosphatidylinositols , Stomach NeoplasmsABSTRACT
Objective To investigate the influence of pertussis toxin(PTX)on G protein-coupled estrogen receptor(GPER)-mediated activation of phosphatidylinositol 3-kinase(PI3K)/protein kinase B (Akt)signaling activated by 17 β-estradiol(17β-E2)in endometrial carcinoma cells.Methods Expressions of GPER protein were detected by immunohistochemical SP method in Ishikawa and HEC-1A cells.Changes of levels of GPER,ERα and ERβ protein and the activation of Akt protein were observed by western blot in the two cells after they were treated by PTX for 30 minutes at different concentrations(0,0.1,0.5,1.0 μg/ml),and then co-stimulated with with 1 × 10-6 mol/L 17β-E2 respectively at different time (Ishikawa 30 minutes,HEC-1A 15 minutes).Results(1)Immunohistochemical SP method showed that GPER was positive stained in cell cytoplasm of Ishikawa and HEC-1A cell.(2)After co-treated with PTX at different concentrations(0,O.1,0.5,1.0 μg/ml)and 10-6 mol/L 17β-E2,in Ishikawa cell,the ratio of pAkt/Akt was 0.74 ±0.54,0.34 ±0.06,0.18 ±0.03,0.07 ±0.15,the gray values of GPER was 0.872 ± 0.490,0.395 ± 0.054,0.145 ± 0.014,0.034 ± 0.008,and with increasing concentration of PTX,the ratio of p-Akt/Akt and the expression of GPER decreased gradually(P ＜ 0.05),which was most obviously when the concentration was 1.0 μg/ml(F =63.729,P =0.0001;F =160.284,P =0.0001);ERα and ERβ protein had no significant change among different groups(P ＞0.05).In HEC-1A cell,the ratio of pAkt/Akt was 0.73 ±0.09,0.26 ±0.14,0.11 ±0.03,0,the Gray values of GPER is 0.927 ±0.134,0.485 ± 0.022,0.194 ± 0.004,0,and with increasing concentration of PTX,the ratio of p-Akt/Akt and the expression of GPER decreased gradually(P ＜ 0.05),which were also completely inhibited when the concentration was 1 μg/ml(F =1039.321,P =0.0001;F =109.646,P =0.0001),ERα protein had no significant differences(P ＞ 0.05)among different groups.ERβ was negatively expressed.Conclusion The results proposed that the activation of PI3K/Akt signaling in Ishikawa and HEC-1A cells could be inhibited after blocking the role of GPER by PTX.
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<p><b>OBJECTIVE</b>To explore the morphology, proliferative activity and adhesion of adipose-derived stem cells (ASC) seeded in the polylactic-co-glycolic acid (PLGA) scaffold, and to provide experimental data for fabricating tissue engineered tympanic membrane repairment materials.</p><p><b>METHODS</b>Wistar rats were selected, and the ASC were isolated and co-cultured with the PLGA scaffold. The morphology and proliferative activity of ASC in the scaffold were examined by immunofluorescence of Vimentin and Ki67 respectively. All the immunofluorescence signals were analyzed by a confocal laser scan microscopy system FLUOVIEW FV1000. The adhesion of ASC to the PLGA scalfold was determined by scanning electron microscopy.</p><p><b>RESULTS</b>Immunofluorescence of vimentin showed rats ASC displayed normal morphology and grew orderly in the PLGA scalfold. Immunofluorescence of Ki67 showed the normal active proliferation of ASC in the scaffolds. The Ki67 mean positive index (x(-) ± s) of the ASC in the scalfold was (8.21 ± 1.76)%, while in control group (cultured without PLGA scalfold) was (9.06 ± 1.95)%. There was no statistic significance between the two groups (t = 1.03, P = 0.30). Scanning electron microscopy demonstrated ASC adhered well to the PLGA scalfold, the pseudopodia of ASC could also be observed and the proliferative cell conjunction was tight.</p><p><b>CONCLUSION</b>ASC has good biocompatible to the PLGA scaffold and could normally adhere and proliferate in PLGA scaffold.</p>
Subject(s)
Animals , Male , Rats , Adipocytes , Cell Biology , Biocompatible Materials , Cell Differentiation , Cells, Cultured , Materials Testing , Rats, Wistar , Stem Cells , Cell Biology , Tissue Engineering , Methods , Tympanic MembraneABSTRACT
<p><b>OBJECTIVE</b>To study the feasibility of applying the Bron-Kerbosch (BK) algorithm on the discovery of basic formulas (BFs) of traditional Chinese medicine.</p><p><b>METHOD</b>This essay introduces the BK algorithm for the discovery of BFs of traditional Chinese medicine and relevant indicators. On the basis of the compatibility network of drugs, with the confidence coefficient of basic formulas anti the support degree to the decline of alpha level as indicators, the BK algorithm was used for the discovery of BFs of traditional Chinese medicine. And prescriptions of Professor Ma Shaoyao, a traditional Chinese medicine (TCM) dermatologist of Longhua hospital, were taken as examples for analysis.</p><p><b>RESULT</b>After the parameters were optimized, three BFs for psoriasis and four BFs for eczema were found using the BK algorithm, which had a relatively high confidence coefficient and basically conformed to the clinical practices.</p><p><b>CONCLUSION</b>The BK algorithm can be used for the discovery of basic formulas (BF) of traditional Chinese medicine, and therefore it is one of the effective methods to study and summarize the diagnosis and treatment experiences and prescription thoughts of famous and experienced TCM doctors.</p>
Subject(s)
Algorithms , Chemistry, Pharmaceutical , Medicine, Chinese TraditionalABSTRACT
<p><b>BACKGROUND</b>Primary malignant germ cell tumors (GCTs) of mediastinum are rare neoplasms. We introduce our institutional experience in managing patients with primary malignant GCTs of the mediastinum, focusing on the analysis of therapeutic modalities.</p><p><b>METHODS</b>A retrospective review was done in 39 consecutive patients with mediastinal malignant GCTs treated in our institution between 1991 and 2007.</p><p><b>RESULTS</b>A total of 39 patients were enrolled in this study with a median age of 27 years. The 5-year overall survival (OS) and progression-free survival (PFS) rates of the whole population were 60.2% and 57.7%, respectively. Stratified by the histology, 18 patients (46.2%) had seminoma and 21 patients (53.8%) had nonseminomatous germ cell tumors (NSGCTs). The 5-year OS rate of patients with seminoma was 87.4% as compared with 36.7% in patients with NSGCTs (P = 0.0004). The 5-year PFS rate was also significantly higher in seminoma patients (87.4% vs. 31.6%, P = 0.003). For 19 patients with NSGCTs managed with multi-modality treatment, chemotherapy exposure appeared to impact the prognosis. The 5-year OS rate was 44.9% in patients with chemotherapy exposure as compared with 20.0% in patients without it (P = 0.43).</p><p><b>CONCLUSION</b>Our study confirmed the significance of systemic chemotherapy in the treatment of primary mediastinal GCTs.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Male , Middle Aged , Young Adult , Disease-Free Survival , Mediastinal Neoplasms , Drug Therapy , Radiotherapy , Neoplasms, Germ Cell and Embryonal , Drug Therapy , Radiotherapy , Retrospective Studies , Seminoma , Drug Therapy , Radiotherapy , Testicular NeoplasmsABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of phosphatidylinositol 3-kinase inhibitor LY294002 combined with NF-κB P65 nuclear translocation inhibitor SN50 on the tumor cell growth and apoptosis using a nude mouse model of gastric cancer.</p><p><b>METHODS</b>Human gastric cancer cell strain SGC7901 was transplanted subcutaneously to nude mice to establish tumor models. Model mice were randomly divided into the control group, the LY294002 treatment group, the SN50 treatment group, and the LY294002+SN50 treatment group, with 5 in each group. After being treated for 10 days, the inhibition rate of tumor growth was ascertained by measuring the size of tumor. Immunohistochemical method was used to detect the expression levels of Bcl-2, P53 and Bax proteins and transmission electron microscopy to investigate the apoptosis of tumor cells.</p><p><b>RESULTS</b>On the 10th day after treatment, the inhibition rate of gastric cancer cellular growth in the LY294002+SN50 group was (49.2±2.5)%, which was significantly higher than that in the LY294002 group(29.4±1.5)% and SN50 group (19.7±1.6)%(P<0.05). In comparison with the other two groups, LY294002+SN50 group exhibited more severe apoptosis, with expression of Bcl-2 decreased and that of P53 and Bax increased more significantly(P<0.05).</p><p><b>CONCLUSION</b>LY294002 combined with SN50 inhibits the growth of SGC7901 transplanted tumor and aggravates the apoptosis of gastric cancer cells in nude mice model.</p>