Efficacy and Safety of Finerenone for Prevention of Cardiovascular Events in Type 2 Diabetes Mellitus With Chronic Kidney Disease: A Meta-analysis of Randomized Controlled Trials.

ABSTRACT: Only a few meta-analyses evaluated the effect of finerenone on cardiovascular events in type 2 diabetes mellitus with chronic kidney disease. The main aim of this meta-analysis was to gain more reliable assessments of the efficacy and safety of finerenone for prevention of cardiovascular events in diabetic kidney disease. We searched for finerenone in the treatment of diabetic kidney disease from database (PubMed, Embase, and ClinicalTrials.gov ) until December 30, 2021. Relative risks (RRs) with 95% confidence intervals (CIs) calculated by the Mantel-Haenszel random-effects model were used as summary statistics for the categorical data. We included 4 studies that met the inclusion criteria with 13,943 participants. The finerenone group demonstrated a great benefit in reducing the incidence of major adverse cardiac events (RR: 0.88; 95% CI 0.80-0.96; P = 0.003), all-cause mortality (RR: 0.89; 95% CI 0.80-0.99; P = 0.04), myocardial infarction (RR: 0.79; 95% CI 0.67-0.92; P = 0.003), and new-onset hypertension (RR: 0.71; 95% CI 0.62-0.81; P < 0.00001). No difference was found in adverse events between the finerenone and placebo groups (RR: 1.00; 95% CI [0.98-1.01], P = 0.59), whereas a higher risk of hyperkalemia was observed in the finerenone group than in the placebo group (RR = 2.04, 95% CI 1.80-2.32; P < 0.00001). Besides, cerebrovascular events and new-onset atrial fibrillation did not increase in patients taking finerenone. Overall, finerenone treatment showed a great benefit of reducing the risk of major adverse cardiac events, all-cause mortality, myocardial infarction, and new-onset hypertension events in patients with type 2 diabetes mellitus and chronic kidney disease.

STAT3-EphA7 axis contributes to the progression of esophageal squamous cell carcinoma.

BACKGROUND: Our previous study has revealed that EphA7 was upregulated in patient-derived esophageal squamous cell carcinoma (ESCC) xenografts with hyper-activated STAT3, but its mechanism was still unclear. MATERIALS AND METHODS: To assess the association between EphA7 and STAT3, western blotting, immunofluorescence, ChIP assay, and qRT-PCR were conducted. Truncated mutation and luciferase assay were performed to examine the promoter activity of EphA7. CCK-8 assay and colony formation were performed to assess the proliferation of ESCC. Cell-derived xenograft models were established to evaluate the effects of EphA7 on ESCC tumor growth. RNA-seq analyses were used to assess the effects of EphA7 on related signals. RESULTS: In this study, EphA7 was found upregulated in ESCC cell lines with high STAT3 activation, and immunofluorescence also showed that EphA7 was co-localized with phospho-STAT3 in ESCC cells. Interestingly, suppressing STAT3 activation by the STAT3 inhibitor Stattic markedly inhibited the protein expression of EphA7 in ESCC cells, in contrast, activation of STAT3 by IL-6 obviously upregulated the protein expression of EphA7. Moreover, the transcription of EphA7 was also mediated by the activation of STAT3 in ESCC cells, and the -2000∼-1500 region was identified as the key promoter of EphA7. Our results also indicated that EphA7 enhanced the cell proliferation of ESCC, and silence of EphA7 significantly suppressed ESCC tumor growth. Moreover, EphA7 silence markedly abolished STAT3 activation-derived cell proliferation of ESCC. Additionally, RNA-seq analyses indicated that several tumor-related signaling pathways were significantly changed after EphA7 downregulation in ESCC cells. CONCLUSION: Our results showed that the transcriptional expression of EphA7 was increased by activated STAT3, and the STAT3 signaling may act through EphA7 to promote the development of ESCC.

Effect of PCSK9 Monoclonal Antibody Versus Placebo/Ezetimibe on Atrial Fibrillation in Patients at High Cardiovascular Risk: A Meta-Analysis of 26 Randomized Controlled Trials.

BACKGROUND: Patients at high cardiovascular risk are closely associated with an increased risk of atrial fibrillation (AF). Whether proprotein convertase subtilisin/kexin type 9 monoclonal antibodies (PCSK9 mAbs) can attenuate AF progression remains unknown. METHODS: To compare PCSK9 mAbs with placebo or ezetimibe to explore the effect of PCSK9 mAbs therapy on the end-point of incidence of AF, we searched PubMed, Embase, and ClinicalTrials.gov for articles. We used Mantel-Haenszel risk ratio (RR) with corresponding 95% CI for the categorical data, including the incidence of AF and predefined other outcomes of interest. RESULTS: We included 21 articles consisting of 26 randomized controlled trials with a total of 95,635 participants. Quantitative synthesis revealed that PCSK9 mAbs significantly reduce the incidence of AF events (RR 0.84; 95% CI 0.72-0.98; p = 0.03), whereas no obvious differences were seen between the PCSK9 mAbs group and the ezetimibe group (RR 0.90; 95% CI 0.29-2.76; p = 0.85). PCSK9 mAbs also markedly decreased the incidence of cerebrovascular events (RR 0.75; 95% CI 0.66-0.85; p < 0.0001) and new-onset hypertension (RR 0.92; 95% CI 0.87-0.97; p = 0.003), but not the risk of cardiovascular death (RR 0.95; 95% CI 0.85-1.07; p = 0.40) and new-onset diabetes mellitus (RR 1.01; 95% CI 0.95-1.08; p = 0.67). CONCLUSIONS: Overall, the PCSK9 mAbs therapy reduced AF and presented certain cardiovascular benefits in patients at high cardiovascular risk. Further big-scale and long follow-up duration randomized controlled trials that compare PCSK9 mAbs with ezetimibe are required to evaluate the effect of PCSK9 mAbs versus ezetimibe on AF.

De-ubiquitination of p300 by USP12 Critically Enhances METTL3 Expression and Ang II-induced cardiac hypertrophy.

Stresses, such as neurohumoral activation, induced pathological cardiac hypertrophy is the main risk factor for heart failure. The ubiquitin-proteasome system (UPS) plays a key role in maintaining protein homeostasis and cardiac function. However, research on the role and mechanism of deubiquitinating enzymes (DUBs) in cardiac hypertrophy is limited. Here, we observe that the deubiquitinating enzyme ubiquitin-specific protease 12(USP12) is upregulated in Ang II-induced hypertrophic hearts and primary neonatal rat cardiomyocytes (NRCMs). Inhibition of USP12 ameliorate Ang II-induced myocardial hypertrophy, while overexpression of USP12 have the opposite effect. USP12 deficiency also significantly attenuate the phenotype of Ang II-induced cardiac hypertrophy in vivo. Moreover, we demonstrate that USP12 aggravate Ang II-induced cardiac hypertrophy by enhancing METTL3, a methyltransferase which catalyze N6-methyladenosine (m6A) modification on messenger RNA and acts as a harmful factor in pathological cardiac hypertrophy. Upregulation of METTL3 reverse the reduction of myocardial hypertrophy induced by USP12 silencing in NRCMs. In contrast, knockdown of METTL3 attenuate the aggravation of myocardial hypertrophy in USP12-overexpressing NRCMs. Furthermore, we discover that USP12 promote the expression of METTL3 via upregulating p300. Mechanistically, USP12 binds and stabilizes p300, thereby activating the transcription of its downstream gene METTL3. Finally, our data show that USP12 is partially dependent on the stabilization of p300 to activate METTL3 expression and promote myocardial hypertrophy. Taken together, our results demonstrate that USP12 acts as a pro-hypertrophic deubiquitinating enzyme via enhancing p300/METTL3 axis, indicating that targeting USP12 could be a potential treatment strategy for pathological cardiac hypertrophy.

[Composition changes reveal relationship between color and enzymatic reaction of Magnoliae Officinalis Cortex during "sweating" process].

In this study, we employed Q Exactive to determine the main differential metabolites of Magnoliae Officinalis Cortex du-ring the "sweating" process. Further, we quantified the color parameters and determined the activities of polyphenol oxidase(PPO), peroxidase(POD), and tyrosinase of Magnoliae Officinalis Cortex during the "sweating" process. Gray correlation analysis was performed for the color, chemical composition, and enzyme activity to reveal the effect of enzymatic reaction on the color of Magnoliae Officinalis Cortex during the "sweating" process. Magnoliae Officinalis Cortex sweating in different manners showed similar metabolite changes. The primary metabolites that changed significantly included amino acids, nucleotides, and sugars, and the secondary metabolites with significant changes were phenols and phenylpropanoids. Despite the different sweating methods, eleven compounds were commonly up-regulated, including L-glutamic acid, acetylarginine, hypoxanthine, and xanthine; six compounds were commonly down-re-gulated, including L-arginine, L-aspartic acid, and phenylalanine. The brightness value(L~*), red-green value(a~*), and yellow-blue value(b~*) of Magnoliae Officinalis Cortex kept decreasing during the "sweating" process. The changes in the activities of PPO and POD during sweating were consistent with those in the color parameter values. The gray correlation analysis demonstrated that the main differential metabolites such as amino acids and phenols were closely related to the color parameters L~*, a~* and b~*; POD was correlated with amino acids and phenols; PPO had strong correlation with phenols. The results indicated that the color change of Magnoliae Officinalis Cortex during "sweating" was closely related to the reactions of enzymes dominated by PPO and POD. The study analyzed the correlations among the main differential metabolites, color parameters, and enzyme activities of Magnoliae Officinalis Cortex in the "sweating" process. It reveals the common law of material changes and ascertains the relationship between color changes and enzymatic reactions of Magnoliae Officinalis Cortex during "sweating". Therefore, this study provides a reference for studying the "sweating" mechanism of Magnoliae Officinalis Cortex and is of great significance to guarantee the quality of Magnoliae Officinalis Cortex.

[Research on network pharmacology of Mongolian medicine Cymbaria in treatment of type 2 diabetes].

The network pharmacology was used to explore the potential active ingredients and action mechanisms of Mongolian medicine Cymbaria in the treatment of type 2 diabetes. According to the literatures collected, Cymbaria component database was established to define important active ingredients and key targets for the anti-hyperglycemic effect to predict action mechanism by active ingredient screening and target prediction techniques. Molecular docking predicted binding activity of main active components with key targets in Cymbaria, then verified the action mechanism in vitro. The Cymbaria component database contained 177 chemical components, 90 chemical structures were confirmed, including 34 chemical components with effective targets. According to the prediction results from network pharmacology, 61 biological processes were significantly affected, such as fatty acid metabolism including PPARs signaling pathway, protein kinase activity and insulin signal pathway. Moreover, the key target proteins were Akt1 and TNFα and quercetin, luteolin and catalpol were the main active ingredients of Cymbaria. Molecular docking prediction showed that luteolin, quercetin and catalpol had a strong binding activity with Akt1; luteolin had strong binding activity but quercetin and catalpol had a certain binding activity with TNFα. Furthermore, catalpol showed hypoglycemic effects in vitro, which up-regulated p-Akt(Ser473)/Akt, PPARα and PPARδ levels and reduced FABP4 expression to regulate glycose and lipid metabolism for improving insulin sensitivity. The network pharmacology predicted that the hypoglycemic effect of Cymbaria was mainly related to anti-inflammatory and lipid regulation with a multi-component, multi-target manner. It provided a scientific view of hypoglycemic effect and action mechanism of Cymbaria for further study.

[Comparative transcriptome and proteome profiling of chlorophyll metabolism pathway in four types of Magnolia officinalis].

Magnolia officinalis is a traditional Chinese medicine,with many years of cultivating process, M. officinalis leaves show more differentiation types due to the exchange of seeds from different provenances. "Da Ao"(DA), "Xiao Ao"(XA), "Chuan Hou"(CH),and "Liu Ye"(LY)are the main types of M. officinalis in Sichuan province of China,and there were obvious differences in growth rate,chemical composition,leaf shape and leaf colour. This study selected different types of M. officinalis leaves(DA,XA,LY and CH)from Sichuan to determine their chlorophyll content. Transcriptomic level sequencing of different types of M. officinalis leaf tissues was by high-throughput sequencing analysis and proteomics used an integrated approach involving TMT labelling and LC-MS/MS to quantify the dynamic changes of the whole proteome of M. officinalis. The results showed that CH had the lowest chlorophyll content while DA had the highest chlorophyll content. Furthermore,transcriptome and proteomics results showed that chlorophyll synthesis pathway in DA glutamine-tRNA reductase,urinary porphyrins decarboxylase(UROD),oxygen-dependent protoporphyrin(ODCO),the original-Ⅲ oxidase protoporphyrin oxidase(PPO),magnesium chelating enzyme subunit ChlD,protoporphyrin magnesium Ⅸ monomethyl ester [oxidative] cyclase(MPPMC)were significantly higher than CH,XA and LY,consistent in the results of determination of chlorophyll content(chlorophyll content was highest of 37.56 mg·g~(-1) FW). Some rate-limiting enzymes related to the chlorophyll synthesis,such as ODCO,PPO and MPPMC were tested by Parallel Reaction Monitoring(PRM),and the results showed that the rate-limiting enzyme content in DA was higher than that in other three types. Therefore,based on the differences in leaf color of four types of M. officinalis,the research conducted a preliminary study on the chlorophyll metabolism pathway in leaves of different types of M. officinalis,and explored relevant genes and proteins causing leaf color differences from the molecular level,so as to lay a foundation for studying the differences in growth and development of different types of M. officinalis.

[Study on regulation of NLRP3/SOCS3-TLR4-NF-κB inflammatory pathway by wogonoside to improve hepatic insulin resistance].

This study was to investigate the hypoglycemic effect of wogonoside to improve hepatic insulin resistance( IR) and its relative anti-inflammatory mechanism. The stable IR-Hep G2 cell model was established by the combination of 1×10-9 mol·L-1 insulin and 3. 75×10-6 mol·L-1 dexamethasone for 48 hours. The changes of glucose consumption in IR-Hep G2 cells with different concentrations of wogonoside( 1,5,10,20,50 µmol·L-1) at different time points( 30,36,48,54 h) were detected by glucose oxidase assay to determine the optimal onset time. Glycogen content and cell viability were respectively detected by ketone method and CCK-8 method. Cryptothermal protein 3( NLRP3),suppressor of cytokine signaling 3( SOCS3),Toll-like receptor 4( TLR4),nuclear factor( NF-κB),interleukin( IL-1ß),IL-6,tumor necrosis factor( TNF-α) involving in the inflammatory signaling pathway,as well as leptin,Ob-R,p-IRS2/IRS2,p-PI3 K/PI3 K( p85),p-Akt/Akt and glucose transporter( GLUT1/2/4) involving in the insulin signaling pathway were detected in IR-HepG2 cells by Western blot. RESULTS: showed that 20 and 50 µmol·L-1 wogonoside significantly up-regulated the glucose consumption of IR-HepG2 cells( P<0. 001) as compared with IR model group,and the optimal onset time was 48 h.Wogonoside had no obvious effect on the cell viability of Hep G2 cells. Further studies showed that 20,50 µmol·L-1 wogonoside respectively increased the glycogen content of IR-HepG2 cells after 48 h treatment,especially in 50 µmol·L-1 group( P<0. 001). Compared with IR model group,wogonoside not only inhibited the protein expression of inflammatory nuclear transcriptional factors NLRP3,SOCS3,TLR4,NF-κB,but also decreased the expression of downstream inflammatory effect factors IL-1ß,IL-6 and TNF-α． In addition,wogonoside elevated Ob-R,p-IRS2/IRS2,p-PI3 K/PI3 K( p85),p-Akt/Akt and GLUT1/2/4 protein expression,whereas it suppressed leptin expression that was regulated by SOCS3. Wogonoside could promote glucose uptake and increase glycogen content to enhance insulin sensitivity in IR-Hep G2 cells. The hypoglycemic effect may be related to the intervention of NLRP3/SOCS3-TLR4-NF-κB inflammatory pathway and decrease of inflammatory factor expression.

[Compound traditional Chinese medicine in treatment of diabetes].

Diabetes is a chronic disorder of glucose metabolism characterized by elevated blood glucose levels. With the improvement in living standards and the changes in lifestyle,T2 DM incidence has a dramatic increase in the past decades,bringing a series of public health problems. In the research and development field of diabetic drugs,T1 DM drugs are mainly long-acting sustained-release insulin preparations,whereas T2 DM drugs mainly are based on single target. T2 DM drugs usually have a good anti-hyperglycemic effect,but with side effects for long-term administration. Therefore,the research and development of hypoglycemic drugs focus on formula drugs with multi-component,multi-target and multi-pathway effects. In the similar principle of action,traditional Chinese medicine formula has achieved a good efficacy in the treatment of diabetes,with mild anti-hyperglycemic effects and multiple-component synergistic effects in intervening the pathogenesis of diabetes,including additive effect,synergy effect and toxicity scattering effect. This article mainly reviews clinical trials and mechanisms of action of traditional Chinese medicine formula for the treatment of diabetes,so as to provide a reference to the rational and effective clinical application of traditional Chinese medicine formula in treating diabetes.

[Microbial community diversity and its characteristics in Magnolia Officinalis Cortex "sweating" process based on high-throughput sequencing].

Magnolia Officinalis Cortex has been used as a traditional Chinese herb for thousands of years in China. According to Chinese Pharmacopoeia,the processing of Magnolia Officinalis Cortex needs " sweating" or " Fahan",which was a special drying process and considered to be an important symbol for high quality and genuine medicinal materials. In this unique processing mode,Magnolia Officinalis Cortex's microbial community structure may be changed,but little is known about microbial diversity during the " sweating". In this study,to analyze the change and its change rules of microbial community of Magnolia Officinalis Cortex in the whole process of " sweating",and find out the microbial community that affects the quality of Magnolia Officinalis Cortex in the process of its " sweating",and provide a basis for further research on the microbial transformation of Magnolia Officinalis Cortex,MiSeq highthroughput sequencing was used to evaluate the microbial diversity of natural " sweating" of Magnolia Officinalis Cortex. In this research,334 genera fungi and 674 genera bacteria were identified. The dominant species weren' t obvious during the early stage of " sweating". Candida was the dominant fungal species( 45. 01%-71. 93%) during the medium " sweating" stage. Aspergillus is the dominant fungal species( 45. 83%-95. 51%) during the late stage of " sweating". Moreover,Enterobacter and Klebsiella were the primary bacterial genus( ≥56. 05%) during the middle and late stages of " sweating". In addition,the predominant bacteria in the process of " sweating" included Bacillus,Deinococcus,Sphingomonas,Hymenobacter and Jatrophihabitans. In conclusion,the microbial diversities and the main dominant fungi and bacteria in the process of " sweating" of Magnolia Officinalis Cortex were initially determined. It was also found that the metabolism of Aspergillus and Candida may be related to the character formation,which were sweet odor and brown inner surface after " sweating". The results provide a theoretical basis for the study of the influence of different microorganisms on the excellent traits formation of " sweating" Magnolia Officinalis Cortex.

[Effect of exogenous GA3 and DPC on active components of Lonicera japonica and its mechanism].

To reveal the effect of plant growth regulator GA3 and DPC on the active components and its possible mechanism of Lonicera japonica. GA and DPC were applied at the stage of flower bud differentiation, and the content of active ingredients was measured by LC-MS-MS, the content of endogenous hormones were measured by ELISA, and the expression of key enzyme enes expression was determined by qRT-PCR. The level of endogenous hormone GA3, IAA, ZR, DHZR and iPA in the GA treatment group was significantly improved, the expression of C4H1, C4H2, 4CL1 and HQT2 were also significantly increased. The content of chlorogenic acid, luteolin, luteoloside, isoquercetin and caffeic acid increased significantly. Spraying DPC did not affect or inhibit the accumulation of active components of L. japonica. Spraying GA can increase the content of endogenous GA3, thus enhance the expression of C4H1, C4H2, 4CL1 and HQT2, and then increase the content of chlorogenic acid and luteolin.

Inhibition of the IGF signaling pathway reverses cisplatin resistance in ovarian cancer cells.

BACKGROUND: This study was aimed at investigating whether metformin can reverse the resistance of ovarian cancer cells to cisplatin and exploring the underlying mechanism. METHODS: Ovarian cancer cell proliferation in vitro was evaluated using a CCK-8 assay. The resistance index of platinum-resistant ovarian cancer cells was determined and cell cycle and apoptosis rate determined by annexin V/propidium iodide double-staining in CP70 cells. Western blotting was used to determine IGF1, IGF1R, AKT, p-IGF1, p-IGF1R, p-AKT, and MRP2 levels in cells treated with different concentrations of metformin and LY29400, an inhibitor of the insulin-like growth factor pathway. Changes in gene expression levels of MRP2, IGF1, IGF1R, and AKT were determined by fluorescence real-time quantitative PCR assay of CP70 cells treated with metformin. Tumors of human ovarian cancer cell lines CP70 and A2780 were established by subcutaneous transplantation of cells in nude mice and the effect of metformin on MRP2 expression and tumor inhibition assessed. RESULTS: The IC50 value of cisplatin in CP70 cells decreased significantly as metformin concentration increased (P < 0.05). The cell cycle distribution in CP70 cells changed with metformin treatment; the percentage of cells in the G0/G1 phase, as well as the natural apoptosis rate was significantly increased with metformin treatment (P < 0.05). IGF1, IGF1R, AKT p-IGF1, p-IGF1R, and p-Akt protein expression was enhanced dose-dependently with metformin, and was also significantly changed by treatment of CP70 cells with 0 mM metformin +10 mM LY294002. Moreover, changes in the expression of MRP2, IGF1, IGF1R, and AKT was metformin-concentration dependent, and was significantly different from that in the untreated control group (P < 0.05). In nude mice, the tumor volumes of the cisplatin-treated groups were significantly less than in the control group, and was further suppressed by co-treatment with cisplatin and metformin (P < 0.05), indicating that these 2 drugs had a synergistic effect on tumor inhibition. CONCLUSION: Metformin can improve the sensitivity of ovarian cancer CP70 cells to cisplatin in a concentration-dependent manner by activating the AKT signaling pathway, inhibiting the IGF1R signaling pathway, and reducing MRP2 expression.

Copper-Promoted Synthesis of 2-Fulleropyrrolines via Heteroannulation of [60]Fullerene with α-Amino Ketones.

A Cu(OAc)2-promoted heteroannulation of [60]fullerene with α-amino ketones has been exploited for the efficient synthesis of 2-fulleropyrrolines containing a trisubstituted or tetrasubstituted C═C bond via the formation of C-C and C-N bonds. Mechanistic study indicates that a radical process should be involved in this transformation. Furthermore, theoretical computations show that the process via the attack of the carbon radical generated from the employed α-amino ketone to [60]fullerene should be the preferred pathway. The electrochemical properties of the synthesized 2-fulleropyrrolines have also been investigated.

Open and Endovascular Management of Severe Cerebral Ischemia in Takayasu's Arteritis.

BACKGROUND: Severe cerebral ischemia in patients with Takayasu's arteries was caused by occlusion of most supra-aortic arteries. Arterial revascularization is necessary to decrease the incidence of stroke and improve the quality of life but may be complicated with multiple occlusive lesions and inflammation condition of this disease. This study was to assess options and long-term outcomes of surgical and endovascular treatment. METHODS: Twenty-nine patients with severe cerebral ischemic symptoms underwent surgical or endovascular treatment from January 1991 to July 2015. Demographic characteristics, surgical and endovascular procedures, and follow-up outcomes were reviewed. Risk factors associated with primary patency of surgical treatment and assisted primary patency of endovascular treatment was identified by Cox regression analyses. RESULTS: There were 29 patients with a median age of 24 (range 9-37 years), 9 in active and 20 in inactive phase. Seventeen patients underwent a variety of bypass procedures. Fourteen endovascular procedures were performed in 12 patients. No death occurred within 30 days after both procedures. Complications within 30 days after bypass included stroke in 1 patient, infection in 2 patients, and heart failure in 1 patient. Nine patients developed brain hyperperfusion after bypass. Transient hemiplegic paralysis occurred in 1 patient during dilation of the carotid artery. During a median follow-up time of 41 months, primary and secondary patency rate of bypass at 1 and 3 years was 93.75% and 100% and 87.5% and 100%, respectively. Assisted primary and secondary patency rate of endovascular treatment at 1 and 3 years was 85.71% and 92.86% and 68.18% and 75.66%, respectively. There was no independent risk factor associated with either primary patency of surgical treatment or assisted primary patency of endovascular treatment. Disease activity was independent risk factor associated with combined rate of primary patency of surgical treatment and assisted primary patency of endovascular treatment (HR: 0.17, 95% CI: 0.03-0.93, P = 0.04). CONCLUSIONS: Bypass is the preferred treatment in majority of patients with good long-term patency, even has a higher propensity for postoperative complications. Endovascular treatment should be preserved for short lesions in inappropriate or high-risk surgical patients but needs more reintervention and close monitoring of lesion for better outcomes. Long-term patency of surgical and endovascular treatment is related with disease activity. Combination of surgical or endovascular treatment and medical therapy may improve the efficacy of interventions.

[Safflower's origin and changes of producing areas].

Human's application of safflower (Carthamus tinctorius) has a long history, but the origin remains unclear. Safflower was introduced into China for traditional Chinese medicine, and Sichuan was major producing area. However, in recent years, the main producing area is in Xinjiang province, in contrast Sichuan safflower is difficult to find. By reading relevant document literature and the production and marketing information of safflower, and having field investigation in the main producing areas, the origin of safflower and the reasons of producing areas' changes were explored. The origin of safflower is considered as the Fertile Crescent in reasonably. The change of producing areas in China is effected by the factors of natural environment and society. The suitability of producing areas and quality of safflower still need to study further.

[Correlation between accumulation of flavoneand expression of functional genes in Carthamus tinctorius].

The average yield of safflower blooming from 1 to 7 day was recorded and calculated, HPLC was used to detect the percentage composition of HYSA,quercetin,naringenin and kaempferol, and the real-time PCR was used to analyze the expression of chs and chi. The average yield,percentage composition of HYSA and naringenin as well as functional genes' expression presented similar trends. The average yield reached the highest peak at the third day, showing highpositive correlation with the contents of HYSA (r=0.756,P<0.05), and significant correlation with the expression of chi (r=0.892,P<0.01). The contents of naringenin showed a high positive correlation with the expression of chs(r=0.766,P<0.05). The study provides a theory basis for the composition and regulation mechanism of the flavonoid constituents and lays foundation for molecular mechanisms which lead to the difference of quality in C. tinctorius.

Subcutaneous dissection area contributes less to endoscopic thyroidectomy-related invasiveness.

OBJECTIVE: To clarify the contribution of the subcutaneous area during breast approach endoscopic thyroidectomy (BAET), with regard to invasiveness-related outcomes. METHODS: Seventy-two patients were randomly assigned to two groups: standard dissection and limited dissection. Postoperative pain and inflammatory response were compared between groups. RESULTS: The groups were well matched except for subcutaneous dissection area (137.11 ± 21.10 vs. 83.69 ± 12.10 cm(2), p < 0.0001). No significant difference was found with regard to VAS score and postoperative inflammatory response. CONCLUSION: Our RCT indicated that the subcutaneous area plays a less important role with regard to BAET-related postoperative pain.

Mutation analysis and prenatal diagnosis in a Chinese family with succinic semialdehyde dehydrogenase and a systematic review of the literature of reported ALDH5A1 mutations.

AIMS: Succinic semialdehyde dehydrogenase (SSADH) deficiency is a neurometabolic disease in which the degradation of γ-aminobutyric acid (GABA) is impaired. The purpose of this study was to report two novel ALDH5A1 mutations responsible for SSADH deficiency in a Chinese family and the prenatal diagnosis of an at-risk fetus with DNA sequencing. RESULTS: Genetic analysis of ALDH5A1, in a child with SSADH deficiency, parents, and 10 weeks' gestation at-risk fetus and 100 healthy unrelated volunteers, was performed. The coding sequence and the intron/exon junctions of ALDH5A1 were analyzed by bidirectional DNA sequencing. The proband was identified to have a compound heterozygous mutations with c.496T>C (p.W166R) and c.589G>A (p.V197M). Each of his parents carried a deleterious mutation. DNA sequencing of chorionic villus revealed the fetus was a carrier, but not affected, and this was confirmed after birth by genetic analysis of umbilical cord blood and urine organic acid analysis. A study in 2003 described 35 mutations of ALDH5A1 in 54 unrelated families, and the current study and systematic literature review identified nine additional novel mutations in eight unrelated families bringing the total number of unique mutations of ALDH5A1 resulting in SSADH deficiency to 44, and the 44 mutations occur from exon 1 to exon 10. No mutational hotspots or prevalent mutations were observed, and all mutations appeared vital for the function of SSADH. CONCLUSIONS: Two novel ALDH5A1 mutations likely responsible for SSADH deficiency were identified, and DNA sequencing provided an accurate diagnosis for an at-risk fetus whose sibling had SSADH deficiency.

Mycorrhiza alters the profile of root hairs in trifoliate orange.

Root hairs and arbuscular mycorrhiza (AM) coexist in root systems for nutrient and water absorption, but the relation between AM and root hairs is poorly known. A pot study was performed to evaluate the effects of four different AM fungi (AMF), namely, Claroideoglomus etunicatum, Diversispora versiformis, Funneliformis mosseae, and Rhizophagus intraradices on root hair development in trifoliate orange (Poncirus trifoliata) seedlings grown in sand. Mycorrhizal seedlings showed significantly higher root hair density than non-mycorrhizal seedlings, irrespective of AMF species. AMF inoculation generally significantly decreased root hair length in the first- and second-order lateral roots but increased it in the third- and fourth-order lateral roots. AMF colonization induced diverse responses in root hair diameter of different order lateral roots. Considerably greater concentrations of phosphorus (P), nitric oxide (NO), glucose, sucrose, indole-3-acetic acid (IAA), and methyl jasmonate (MeJA) were found in roots of AM seedlings than in non-AM seedlings. Levels of P, NO, carbohydrates, IAA, and MeJA in roots were correlated with AM formation and root hair development. These results suggest that AMF could alter the profile of root hairs in trifoliate orange through modulation of physiological activities. F. mosseae, which had the greatest positive effects, could represent an efficient AM fungus for increasing fruit yields or decreasing fertilizer inputs in citrus production.

[Cloning,Bioinformatic Analysis of Chalcone-Flavonone Isomerase Gene( CHI) and Relationship Between Expression of CHI and Accumulation of HSYA in Carthamus tinctorius].

Objective: To clone chalcone-flavonone isomerase( CHI) gene in Carthamus tinctorius,to analyze the bioinformation of CHI,to compare the expression of CHI,and to analyze the percentage composition of hydroxysafflor yellow A( HSYA) during the florescence,in order to provide the foundation for functional verification of CHI and the composition and regulation mechanism of the flavonoid constituents in Carthamus tinctorius. Methods: CHI was cloned,bioinformatics was used to analyze the protein characteristics, real timePCR was used to analyze the expression of CHI,and HPLC was used to analyze the percentage composition of HSYA Results: A 696 bp CHI sequence in Carthamus tinctorius,the expression of CHI and the percentage composition of HSYA during the florescence were obtained. The variation tendency was similar between the expression of CHI and the accumulation of HSYA, which was increased gradually during 1 ~ 4 d and peaked at the fourth day,then decreased sharply during the fifth to the seventh day of florescence. Conclusion: This research provides a foundation for functional verification of CHI and the composition and regulation mechanism of the flavonoid constituents in Carthamus tinctorius.