Differential metabolite analysis of the pharmacodynamic differences between different ratios of Dahuang ()-Taoren () herb pair.

OBJECTIVE: To explore the material basis of the difference of efficacy of Dahuang (Radix Et Rhizoma Rhei Palmati)-Taoren (Semen Persicae) (DT) drugs with different proportions. METHODS: Samples of different ratios of Dahuang (Radix et Rhizoma Rhei Palnati, DH) to Taoren (Semen Persicae, TR) (Group A 1:1, B 2:3, C 3:2) were analyzed based on gas chromatography time-of-flight mass spectrometry untargeted metabolomics technique. RESULTS: A total of 240 primary metabolites were detected. Forty-one differential metabolites involved nine differential metabolic pathways, of which four were closely related to the efficacy of DT in the treatment of heat and blood stasis syndrome. These pathways included the biosynthesis of amino acid (phenylalanine tyrosine and tryptophan), flavonoids, unsaturated fatty acids, and the glycolysis/glycogenesis pathway. CONCLUSION: There are significant differences in the efficacy of different ratios of DT drugs, and their optimal ratio for the treatment of heat and blood stasis syndrome should be 1:1.

Treatment of Cerebral Ischemia-reperfusion Injury by Traditional Chinese Medicine Based on PI3K/Akt Signaling Pathway： A Review / 中国实验方剂学杂志

Cerebral ischemia/reperfusion injury （CIRI） is a complex cascade reaction process in which the blood flow and oxygen supply of brain tissue in the infarcted area recover after cerebral ischemia， resulting in secondary injury of ischemic brain tissue. At present， thrombolysis as soon as possible and restoration of cerebral blood supply are still the only strategies for the treatment of stroke， but a considerable number of patients' symptoms will be more serious after reperfusion， making patients face adverse outcomes such as neurological function injury and even death and seriously affecting the quality of life and safety of patients. Therefore， an in-depth exploration of the mechanism and treatment strategy of CIRI has important clinical significance. The phosphatidylinositol 3- kinase （PI3K）/protein kinase B （Akt） signaling pathway is one of the classic anti-apoptosis/reproductive-promoting signal transduction pathways， which is responsible for cell proliferation， growth， and differentiation. It is the key cascade signaling pathway of CIRI， located at the core site in many mechanisms such as mitochondrial disorder， apoptosis， autophagy， oxidative stress， and inflammation. It is closely related to the occurrence and development of CIRI. Traditional Chinese medicine has been used in the clinical treatment of stroke and its complications for thousands of years， and the clinical effect of traditional Chinese medicine in the prevention and treatment of CIRI has been affirmed by a large number of research results in recent years. It is further clarified that the monomers， active components， and their compound prescriptions of traditional Chinese medicine can directly or indirectly regulate the PI3K/Akt signaling pathway by virtue of the biological advantages of multi-targets， multi-components， and multi-pathways and play an overall protective role in CIRI. By analyzing the related research progress of traditional Chinese medicine in China and abroad in recent years， the authors summarized the role and mechanism of regulating the PI3K/Akt signaling pathway in the prevention and treatment of CIRI， so as to provide further theoretical basis for the study of the mechanism of clinical prevention and treatment of CIRI.

AP2M1 inhibits proliferation and invasion of diffuse large B-cell lymphoma cells / 基础医学与临床

Objective To evaluate the regulatory effect of the adaptor related protein complex 2 subunit μ1(AP2M1)on proliferation and invasion of diffuse large B-cell lymphoma(DLBCL).Methods Human diffuse large B-cell lymphoma cell line OCI-LY8 was aliquoted into control group,NC-shRNA group,AP2M1-shRNA group,NC-LV group,and AP2M1-LV group.Lipofectamine 2000 was used for cell transfection.Cell proliferation was detected by tetramethylazolium salt(MTT)method,apoptosis was detected by flow cytometry and cell migration and invasion were detected by Transwell assay.The protein expression of AP2M1,epidermal growth factor receptor(EGFR),p-phosphatidylinositol 3 kinase(PI3K),PI3K,p-protein kinase B(Akt)and AKT was detec-ted by Western blot.Results Compared with control group,the relative expression of AP2M1 mRNA and protein in the AP2M1-shRNA group was decreased(P＜0.05).The relative cell viability was increased(P＜0.05).The cell apoptosis rate was decreased(P＜0.05).The counting number of migrating and invading cells was in-creased(P＜0.05).The relative expression level of EGFR protein and the phosphorylation level of PI3K and AKT were increased(P＜0.05).Compared with Control group,the expression of AP2M1 mRNA and protein relative ex-pression level in AP2M1-LV group was increased(P＜0.05).The relative cell viability was decreased(P＜0.05).The cell apoptosis rate was increased(P＜0.05).The number of migrating and invading cells was decreased(P＜0.05).The relative expression level of EGFR protein and the phosphorylation level of PI3K and AKT were all decreased(P＜0.05).Conclusions The over-expression of AP2M1 partially inhibits the proliferation and invasion of DLBCL cells by inhibiting the EGFR/PI3K/AKT signaling pathway.

Expression of lncRNA SNHG8 in placenta accreta and its effect on trophoblast invasion and migration / 实用医学杂志

Objective To investigate the expression of lncRNA SNHG8 in placenta accrete(PA)and its effect on trophoblast invasion and migration.Methods qRT-PCR was used to detect the expression of lncRNA SNHG8 in placenta tissue of 30 cases in PA group and 30 cases in control group,and the correlation between lncRNA SNHG8 expression and prenatal ultrasound score of 30 cases in PA group was analyzed.Transwell and scratch assay were used to detect the effect of lncRNA SNHG8 interference on the invasion and migration of human chorionic trophoblast cells(HTR8/SVneo cells),and western blot was used to detect the expression of MMP-2 and MMP-9.The downstream targets of lncRNA SNHG8 were predicted by StarBase software,and the expression of lncRNA SNHG8 was detected in placental tissues of the two groups.Dual luciferase reporter assay was used to detect the targeting relationship between lncRNA SNHG8 and miR-542-3p.Results Compared with that of the control group,the expression of lncRNA SNHG8 was up-regulated in the placenta tissue of the PA group(P<0.05),and it was positively correlated with prenatal ultrasound score.Interference with lncRNA SNHG8 inhibited the invasion and migration of trophoblast cells(P<0.05);the protein expression of MMP-9 and MMP-2 also decreased signifi-cantly(P<0.05).Biological prediction indicates that miR-542-3p had a binding site with lncRNA SNHG8,and miR-542-3p expression was down-regulated in PA placental tissue(P<0.05).Dual luciferase reporter assay confirmed that lncRNA SNHG8 could target miR-542-3p.Compared with si-SNHG8+inhibitor-NC,co-transfection of si-SNHG8 and miR-542-3p inhibitor enhanced the invasion and migration ability of trophoblast cells(P<0.05).Conclusion lncRNA SNHG8 is highly expressed in PA and is related to the severity of PA.LncRNA SNHG8 promotes the invasion and migration of trophoblast by regulating the level of miR-542-3p.The study suggests that lncRNA SNHG8 plays an important role in the invasion and migration of PA trophoblast cells,which is expected to be a clinical diagnostic biomarker and therapeutic target.

Research progress of the endoplasmic reticulum-mitochondrial interactions in post-stroke cognitive impair-ment / 实用医学杂志

Post-stroke cognitive impairment(PSCI),refers to a range of clinical syndromes of cognitive impairment caused by stroke.Although its specific pathogenesis is still unclear,many studies have confirmed that endoplasmic reticulum-mitochondria interaction has become a key hub for intracellular signal transduction and substance metabolism,and its regulation of various biological processes,such as Ca2+ balance,lipid metabolism,mitochondrial dynamics,autophagy,and neuroinflammation,is closely related to the development of PSCI.There-fore,in this paper,we will review the various functions of endoplasmic reticulum-mitochondrial interactions and explore their specific roles in PSCI,in order to discover new therapeutic targets and provide new theoretical basis and references for the development of PSCI-targeted drugs in the future.

Research progress on the potential mechanism and application of lactobacillus in the prevention and treatment of acute myocardial infarction / 天津医药

With the development of clinical,basic medicine and metabolomics,intestinal flora has gradually become a focus of research and has been used in the diagnosis and treatment of several intestinal diseases.Lactobacillus can produce and regulate a variety of intestinal metabolites,and participate in lipid metabolism,thrombosis,inflammation and oxidative stress,and even ferroptosis,which may have an impact on the occurrence and development of cardiovascular diseases.This article reviews the role of lactobacillus in the regulation of metabolism,involvement in the pathophysiological mechanism of myocardial infarction and its clinical application in acute myocardial infarction,providing a new reference for the prevention and treatment of AMI.

Comparison of clinical outcomes in patients with 4b acute myocardial infarction caused by early and late stent thrombosis / 天津医药

Objective To observe and compare in-hospital and 1-year survival and prognosis of patients with 4b acute myocardial infarction(AMI)caused by early and late stent thrombosis(ST).Methods A total of 302 patients with 4b acute myocardial infarction caused by ST were enrolled in this study from January 2015 to February 2018.ST patients were confirmed by coronary angiography.These patients were divided into two groups:the early ST group(n=26)and the late ST group(n=276)according to the time of ST occurrence.Endpoint events during hospitalization and one year of follow up were compared between the two groups of patients.The primary endpoint events included cardiac death and recurrent AMI.The secondary endpoint events included target lesion revascularization(TLR),re-stent thrombosis,heart failure and stroke.The incidence of no endpoint events was compared between two groups of patients by Kaplan and Meier survival analysis.Cox regression analysis was used to analyze risk factors for endpoint events in patients with type 4b AMI.Results There was no significant difference in the incidence of the primary endpoint events during hospitalization between the two groups(7.7%vs.3.3%,P=0.243).The incidence of heart failure was higher in the early ST group than that of the late ST group(11.5%vs.1.4%,P=0.016).There was no significant difference in the incidence rates of other secondary endpoint events between the two groups(P＞0.05).After a mean follow-up of 1 year,the incidence rates of primary endpoint events and the secondary eendpoint events were higher in the early ST group(20.0%vs.5.9%,P＜0.05 and 36.0%vs.11.5%,P＜0.01)than that of the late ST group.Kaplan and Meier survival analysis showed that the 1-year cumulative incidences of non-primary(P= 0.022)and non-secondary events(P＜0.001)were lower in the early ST group than those of the late ST group.Cox regression analysis showed that hypertension and history of coronary artery bypass grafting were independent risk factors for primary endpoint events in patients with 4b AMI,and intraoperative implantation of intraaortic balloon pump(IABP)and shortening the time from stent thrombosis to balloon dilation(ST to B)were independent protective factor.Conclusion Early ST patients have similar in-hospital outcomes and poor long-term prognosis compared with late ST induced 4b type AMI patients.Intraoperative implantation of IABP and shortening of ST to B time may improve the prognosis of 4b type AMI patients.

Research progress on association between chronic diseases and herpes zoster / 中华全科医师杂志

Herpes zoster is an infectious skin disease caused by reactivation of the varicella-zoster virus with multiple manifestations and various complication clinically. Studies have confirmed that chronic diseases are one of the independent risk factors for HZ; and the common chronic diseases such as diabetes, cardiovascular disease, chronic obstructive pulmonary disease and rheumatoid arthritis are associated with the development of herpes zoster. In this article, we review the latest research progress on the clinical features of herpes zoster, its correlation with common chronic diseases and the prevention strategies to reduce the disease burden.

Impact of smart physical education assignment on physical health of male college students / 中国学校卫生

Objective@#To explore the impact of smart physical education assignment on physical health of male university students, so as to provide theoretical support and practical references for physical health improvement of male university students and implementing smart sports assignments.@*Methods@#From September 2023 to January 2024, 317 sophomore male students from six Taekwondo elective classes at Hunan Institute of Engineering were selected and were randomly divided into an intervention group (n=157) and a control group (n=160). The intervention group was given sports assignments twice a week through smart means with an intervention duration of 15 weeks, each time for 25-35 minutes, in addition to the teaching according to the public course syllabus, while the control group was taught according to the public course syllabus. The physical and health indicators of both groups were tested before and after intervention,then the differences in various physical health indicators between two groups of students before and after intervention were compared through ttest and Mann-Whitney U test.@*Results@#After the intervention, the vital capacity, 50 m run, sitandreach, 1 000 m run, and pullup scores of the intervention group significantly improved compared to those before intervention. The scores improved from (3 918.27±737.34)mL, 7.88(7.53,8.45)s, 9.80(2.70,15.75) cm, 4.30(4.12,4.50) min and 3.00(0.00,7.50) times to (4 574.19±800.61) mL, 7.65(7.37,8.12)s, 17.20(11.80,21.55)cm, 4.13(3.58,4.31)min and 5.00(1.00,10.00) times,respectively (t/Z=-7.60, 2.61, -8.39, 5.62, -2.72, P<0.05). Before intervention, there was no statistically significant difference in physical health indicators between the intervention group and the control group (P>0.05).After intervention,the scores of the intervention group on the vital capacity,50 m run,sitandreach,1 000 m run and pullup, were significantly higher than those of the control group [(4 310.97±808.90)mL, 7.75(7.40,8.30)s, 14.10(8.42,17.87)cm, 4.29(4.08,4.45)min and 4.00(1.00,7.00) times] (t/Z=2.91, -4.55, -4.75, -4.15, 2.58, P<0.05).@*Conclusions@#Having 25-35 min smart physical education assignment twice a week can effectively improve physical health level of male college students. It is recommended to assign appropriate amount of smart sports homework to improve physical health level of college students, while ensuring the amount and intensity of physical activity in public physical education courses.

Moral Dilemma and Research on Coping Style of Frontline Nurses During the Outbreak of COVID -19 / 中国医学伦理学

【Objective:】 To understand the moral dilemma, coping style of frontline nurses and their correlation during the COVID -19. 【Methods:】 From March to April, 2021, 651 nurses working in designated hospitals for COVID -19 were selected by convenient sampling, and the Chinese version of Moral Distress Scale-Revised (MDS-R) and Chinese version of Coping Style Questionnaire (CSQ) were used for investigation. 【Results:】 Frontline nurses’ MDS-R score was 91.0, "value conflicts" was the moral dilemma with the highest frequency and intensity, more than half (51.0%) of the respondents had considered leaving their position because of moral dilemma. "Problem solving" and "seeking for help" were the major coping strategies adopted by the nurses. Nurses who directly provide medical services for COVID -19 patients, dissatisfied with hospital-related training and adopted negative coping strategies had higher moral dilemmas than other nurses. 【Conclusion:】 The moral dilemma of frontline nurses is at a high level during the period of epidemic prevention and control. Nursing managers should pay attention to guiding and helping nurses to establish positive coping styles and improve their ability to respond to major public health emergencies.

Research progress in the route of administration and corresponding dosage form of triamcinolone acetonide / 中国药房

As a glucocorticoid drug with wide clinical application， triamcinolone acetonide can be administered by multiple routes， such as eye， nose， joint cavity， and skin， for the treatment of various local diseases such as arthritis， macular edema， rhinitis， and urticaria. As a drug with extremely low solubility in water， the dose form of triamcinolone acetonide is closely correlated with administration route and site. The dosage form of triamcinolone acetonide administered via injection（including joint cavity injection， vitreous injection， suprachoroidal injection， intramuscular injection） is mainly suspension， and the representative drugs include Kenalog-40®， Zilretta®， Triesence®， Xipere®， etc.； the dosage forms of nasal mucosal administration are mostly sprays， and the representative drug is Nasacort®； the dosage forms of oral mucosal administration are mostly patches， ointments and creams， and the representative drug is Oracort®； the dosage forms for transdermal administration are mostly ointments， creams and lotions， and the representative drugs include Trianex®， Teva-Triacomb®， etc. At present， the research on dosage forms of triamcinolone acetonide by various administration routes mainly focuses on the construction of delivery carriers， the addition of cosolvents or the use of new delivery tools.

Survival rate and quality of life in patients with liver cirrhosis complicated with bacterial infection / 公共卫生与预防医学

Objective To analyze the short-term survival and prognostic quality of life of patients with liver cirrhosis complicated by bacterial infection. Methods This study collected and analyzed 300 patients with liver cirrhosis complicated with infection who were hospitalized in the First Affiliated Hospital of Hebei North University, and followed up to discuss their survival and quality of life. Results In this study, the top two causes of infection were spontaneous bacterial peritonitis (60.67% of patients) and pneumonia (50.67% of patients). The second causes were urinary tract infections (15.33%), gastrointestinal infections (12.33%), and other causes. There was no statistically significant difference between male and female patients (P>0.05). In addition, the proportion of hospital infections was 71.00%, and there was no statistically significant difference between male and female patients (P>0.05). A total of 353 strains of pathogenic bacteria were isolated in this study (73.37% of patients with hospital infections). The distribution analysis of pathogenic bacteria showed that the highest proportion of ECO was 35.98%, followed by Klebsiella pneumoniae (18.98%). The distribution trend of 259 strains of pathogenic bacteria among hospital patients was consistent with that of all strains, and the difference was not statistically significant (P>0.05). Gram negative bacteria accounted for 79.60% (281/353) of all detected strains, of which Escherichia coli was mostly detected in patients with spontaneous bacterial peritonitis, Klebsiella Pneumoniae (KPN) and Pseudomonas aeruginosa (PAE) were mostly detected in patients with pneumonia, and Enterococcus (ENF) was mostly detected in patients with urinary tract infection; Among gram-negative bacteria, Staphylococcus epidermidis (SEP) and Staphylococcus aureus (SAU) are mostly found in patients with other infectious causes (blood flow infection, etc.) , and Streptococcus (STR) accounts for a high proportion in patients with Spontaneous bacterial peritonitis. In this study, 9 cases of death prognosis were detected during follow-up, and there was no statistically significant difference in the detection of death prognosis between different bacterial strains in both genders, as well as the difference in detection of death prognosis between hospital infections and out of hospital infections in both genders (P>0.05). There was no statistically significant difference in the detection of death prognosis between males and females due to different causes of infection, P>0.05. The quality of life scores of 291 surviving patients were compared between baseline and follow-up, indicating an increase in follow-up scores, especially in the dimensions of physiological function and physical pain. There was no statistically significant difference between different bacterial strains, infection causes, and hospital/non hospital infections (P>0.05) . Conclusion Spontaneous bacterial peritonitis and pneumonia are the main causes of infection that deserve special attention, and the main pathogens of infection are Gram negative bacteria. Targeted treatment and rehabilitation should be provided for patients with liver cirrhosis complicated by infection. At the same time, the proportion of hospital infections is relatively high, and attention should be paid to, prevention and control measures should be implemented as well.

Validity and reliability of the Chinese version of the Body Acceptance by Others Scale-2 in adults / 中国心理卫生杂志

Objective:To examine the validity and reliability of the Chinese version of the Body Acceptance by Others Scale-2(BAOS-2)in adults.Methods:Totally 616 adults(aged 18-56 years)were selected to test the structural validity,and internal consistency reliability of the scale.The Body Appreciation Scale-2(BAS-2),Func-tionality Appreciation Scale(FAS),Self-Compassion Scale(SCS),Self-Esteem Scale(SES)and Satisfaction with Life Scale(SWLS)were used as criteria to test criterion validity.A sample of 55 adults was retested 2 weeks later for the test-retest reliability.Results:The exploratory factor analysis extracted one factor,and the factor loading range of each item was 0.59-0.76.The confirmatory factor analysis showed that the factor model fit indices were acceptable(x2/df=2.99,CFI=0.91,GFI=0.90,TLI=0.89,RMR=0.04,RMSEA=0.08).The scores of the Chinese version of BAOS-2 were positively correlated with the scores of BAS-2,FAS,SCS,SES and SWLS(r=0.43-0.66,Ps＜0.001).The Cronbach a of the Chinese version of BAOS-2 was 0.90,and the test-retest reliabili-ty(ICC)was 0.65.Conclusion:The Chinese version of the Body Acceptance by Others Scale-2(BAOS-2)has i-deal validity and reliability.

Phosphatidylinositol 3-kinases Play a Suppressive Role in Electrotaxis of Dictyostelium Cells Through Akt and ERK / 生物化学与生物物理进展

ObjectivePhosphatidylinositol 3 kinases (PI3Ks) play an important role in cell directional movement by regulating F-actin. However, the structure and function of PI3Ks are complex. The role of PI3Ks in cell electrotaxis is not fully understood. Therefore, in this study, the model organism Dictyostelium discoideum cells were used as experimental materials to explore the role of PI3K1 and PI3K2 in electrotaxis. MethodsFirstly, PI3K1 coding gene pikA knockout mutant and PI3K2 coding gene pikB knockout mutant were constructed by CRISPR/Cas9 system. Secondly, two mutants were placed in a DC electric field with a strength of 12 V/cm and the electrotaxis were analyzed. ResultsData analysis showed that the direction index of wild-type cells in DC electric field was (0.86±0.03), while the direction index of pikA- and pikB- mutants in DC electric field was (0.95±0.02) and (0.94±0.03), respectively. In addition, the average trajectory speed of wild-type cells in the electric field was (3.34±0.08) μm/min, while the average trajectory speed of pikA- and pikB- mutants were (4.85±0.20) μm/min and (5.48±0.15) μm/min, respectively. The t test showed that there were significant differences in the directedness index and speed between the mutant and wild type. Western blot results showed that both phosphorylated Akt and phosphorylated ERK were significantly increased in pikA- and pikB- mutants. ConclusionPI3K1 and PI3K2 may inhibit the electrotaxis of Dictyostelium discoideum cells by increasing the activity of Akt and ERK.

Health literacy of chronic diseases in newly diagnosed and prediabetes patients and its relationship with the effect of blood glucose control / 公共卫生与预防医学

Objective To explore the relationship between health literacy of chronic diseases and the effect of blood glucose control in newly diagnosed and prediabetes patients. Methods A total of 180 newly diagnosed and prediabetes patients treated in the Western Theater Command General Hospital from January 2021 to January 2023 were selected, including 94 newly diagnosed and 86 prediabetes patients. The health literacy of chronic diseases in these patients was evaluated by using the health literacy management scale (HeLMS) developed by Jordan et al. The differences of clinical general information and blood glucose control between patients with adequate and insufficient health literacy were analyzed. Results Among the 180 patients, the total score of information acquisition ability, communication and interaction ability, willingness to improve health, willingness for financial support, total score of health literacy, and adequate proportion of health literacy assessed by HeLMS were (35.54 ± 7.21), (33.02 ± 8.15), (15.54 ± 3.92), (7.10 ± 0.98), (91.87 ± 7.28), and 51.67%, respectively. The age of patients with adequate health literacy in the newly diagnosed diabetes group was (52.23 ± 7.15) years old, which was significantly lower than that of patients with insufficient health literacy (P<0.05). The proportions of patients with high school education and above, monthly income ≥ 3000 yuan, and relatives engaged in medical work were 68.09%, 68.09%, and 34.04%, respectively, which were significantly higher than those of patients with insufficient health literacy (P<0.05). The age of patients with adequate health literacy in prediabetes group was (82.23 ± 4.15) years old, which was significantly lower than that of patients with insufficient health literacy (P<0.05). The proportions of patients with high school education and above, monthly income ≥ 3000 yuan, number of children ≥ 2, and relatives engaged in medical work were 65.22%, 67.39%, 34.78%, and 41.30%, respectively, which were significantly higher than those of patients with insufficient health literacy (P<0.05). The fasting blood glucose, glycosylated hemoglobin and 2h postprandial blood glucose of patients with adequate health literacy in the newly diagnosed diabetes group after treatment were (6.43 ± 0.93) mmol/L, (6.02 ± 0.91)% and (7.71 ± 1.01) mmol/L, respectively, which were significantly lower than those of patients with insufficient health literacy (P<0.05). The fasting blood glucose, glycosylated hemoglobin and 2h postprandial blood glucose in patients with adequate health literacy in prediabetes group after treatment were (5.21 ± 0.37) mmol/L, (5.20 ± 0.40)% and (6.20 ± 0.92) mmol/L, respectively, which were significantly lower than those in patients with insufficient health literacy (P<0.05). Conclusion The chronic disease health literacy of prediabetes and new-onset patients is related to the age, education and monthly income level, number of children, relatives engaged in medical work of patients, and the level of health literacy is helpful to the control of blood glucose.

Seven-layer analysis model of an optical waveguide excitation fluorescence microscopy.

In this paper, an optical waveguide evanescent field fluorescence microscopy is studied. Based on Maxwell's equation, a seven-layer theoretical analysis model is developed for the evaluation of an optical waveguide excitation fluorescence microscopy. The optical waveguide excitation fluorescence microscopy structure is systematically and comprehensively analysed at the wavelengths of 488, 532 and 646 nm for fluorescent dyes. The analysis results provide some useful suggestions, which will be beneficial to the research of an optical waveguide evanescent field fluorescence microscopy.

Co-expression of soluble guanylyl cyclase subunits and PDE5A shRNA to elevate cellular cGMP level: A potential gene therapy for myocardial cell death.

BACKGROUND: Genetic manipulation on the NO-sGC-cGMP pathway has been rarely achieved, partially due to complexity of the soluble guanylyl cyclase (sGC) enzyme. OBJECTIVE: We aim to develop gene therapy directly targeting the pathway to circumvent cytotoxicity and tolerance after prolonged use of NO-donors and the insufficiency of PDE inhibitors. METHODS: In this study, we constructed lentivirus vectors expressing GUCY1A3 and GUCY1B3 genes, which encoded the α1 and ß1 subunits of soluble guanylyl cyclase (sGC), respectively, to enhance cGMP synthesis. We also constructed lentiviral vector harboring PDE5A shRNA to alleviate phosphodiesterase activity and cGMP degradation. RESULTS: Transductions of human HEK293 cells with the constructs were successful, as indicated by the fluorescent signal and altered gene expression produced by each vector. Overexpression of GUCY1A3 and GUCY1B3 resulted in increased sGC enzyme activity and elevated cGMP level in the cells. Expression of PDE5A shRNA resulted in decreased PDE5A expression and elevated cGMP level. Co-transduction of the three lentiviral vectors resulted in a more significant elevation of cGMP in HEK293 cells without obvious cytotoxicity. CONCLUSION: To the best of our knowledge, this is the first study to show that co-expression of exogenous subunits of the soluble guanylyl cyclase could form functional enzyme and increase cellular cGMP level in mammalian cells. Simultaneous expression of PDE5A shRNA could alleviate feedback up-regulation on PDE5A caused by cGMP elevation. Further studies are required to evaluate the effects of these constructs in vivo.

Identification and analysis of R1-MYB gene family in Rheum palmatum L. based on full-length transcriptome sequencing / 药学学报

As one kind of v-myb avian myeloblastosis viral oncogene homolog (MYB) transcription factors, R1-MYB (MYB-related) family plays an important role in plant growth and development, as well as environmental stress and hormone signal transduction. In this study, R1-MYB family genes in Rheum palmatum L. were systematically screened based on full-length transcriptome sequencing analysis. Firstly, the physicochemical, protein domain and molecular evolution characteristics of the coding proteins were analyzed. Furthermore, the tissue expression levels of R1-MYB genes were analyzed by RNA-seq. We also investigated the expression pattern of RpMYB24 in response to various hormones and abiotic stresses. The results showed that a total of 49 R1-MYB genes were identified, which mainly encoded thermally stable hydrophilic proteins. Most of the deduced proteins were predicted to locate in nucleus. Each protein had a large proportion of random curl and α helix, and also had the W-type conserved amino acids which were the signature of MYB. R1-MYB family members were distributed in five subgroups, including circadian clock associated 1 (CCA1)-like, I-box (GATAAG)-like, CAPRICE (CPC)-like, telomere repeat binding factor (TRF)-like and TATA binding protein (TBP)-like, and the number of CCA1-like was the majority. RNA-seq revealed that 49 R1-MYB genes were differentially expressed in roots, rhizomes and leaves of R. palmatum, and the expression levels of 15 and 23 genes in roots and rhizomes were higher than those in leaves, respectively. RpMYB24 transcript was induced by abscisic acid (ABA), salicylic acid (SA), and methyl jasmonate (MeJA) treatment, and could also significantly respond to injury, low temperature and high temperature stresses except drought stress. This study systematically identified the R1-MYB family genes and their molecular characteristics, better for further gene functional validation, and then provide a scientific basis for the transcriptional regulation mechanism research into rhubarb quality formation.

Synthesis and anti-HCC activity of full 2ʹ-F/OMe-siRNA encapsulated with neutral cytidinyl/cationic lipid / 药学学报

A variety of full 2ʹ-F/OMe-modified siRNAs were designed and synthesized, and the activity against hepatocellular carcinoma Huh-7 and HepG2 cells was evaluated. K&A DNA/RNA H-8 synthesizer was used to synthesize siRNAs, and neutral cytidinyl lipid DNCA mixed with cationic lipid CLD were used to transfect siRNA. By RT-qPCR and CCK-8 assay, the target gene silence and the proliferation of Huh-7 and HepG2 cells were detected. The siRNAs loading into Ago2 protein was detected by RNA-binding protein immunoprecipitation. Drug uptake and cell apoptosis were detected by flow cytometry, and the expression of PLK1 protein was detected by Western blot. Partial full 2ʹ-F/OMe modified siRNAs, especial siPLK1A3, increased the uptake of Huh-7 cells, enhanced their binding to Ago2 and gene silencing activity, down-regulated PLK1 protein, as well as induced more Huh-7 cell apoptosis and proliferation inhibition activity. It provides important data for the development of novel siRNA modification patterns and anti-HCC formulations.

Variation and interaction mechanism between active components in Rheum officinale and rhizosphere soil microorganisms under drought stress / 中国中药杂志

To explore the changes and the reaction mechanisms between soil microecological environment and the content of secon-dary metabolites of plants under water deficit, this study carried out a pot experiment on the 3-leaf stage seedlings of Rheum officinale to analyze their response mechanism under different drought gradients(normal water supply, mild, moderate, and severe drought). The results indicated that the content of flavonoids, phenols, terpenoids, and alkaloids in the root of R. officinale varied greatly under drought stresses. Under mild drought stress, the content of substances mentioned above was comparatively high, and the content of rutin, emodin, gallic acid, and(+)-catechin hydrate in the root significantly increased. The content of rutin, emodin, and gallic acid under severe drought stress was significantly lower than that under normal water supply. The number of species, Shannon diversity index, richness index, and Simpson index of bacteria in the rhizosphere soil were significantly higher than those in blank soil, and the number of microbial species and richness index decreased significantly with the aggravation of drought stresses. In the context of water deficit, Cyanophyta, Firmicutes, Actinobacteria, Chloroflexi, Gemmatimonadetes, Streptomyces, and Actinomyces were the dominant bacteria in the rhizosphere of R. officinale. The relative content of rutin and emodin in the root of R. officinale was positively correlated with the relative abundance of Cyanophyta and Firmicutes, and the relative content of(+)-catechin hydrate and(-)-epicatechin gallate was positively correlated with the relative abundance of Bacteroidetes and Firmicutes. In conclusion, appropriate drought stress can increase the content of secondary metabolites of R. officinale from physiological induction and the increase in the association with beneficial microbe.