Evaluation of surgical revascularization procedure outcomes for adult Moyamoya disease: a computed tomography perfusion-based study.

BACKGROUND: The effectiveness of surgical interventions, whether direct or indirect, for Moyamoya disease (MMD) remains controversial. This study aims to investigate CT perfusion (CTP) as an objective method to evaluate the outcomes of different surgical modalities for adult MMD. METHODS: The clinical and imaging data of 41 patients who underwent superficial temporal artery-middle cerebral artery (STA-MCA) bypass and 43 who received encephaloduroarteriosynangiosis (EDAS) were retrospectively analyzed. Intra- and intergroup differences in the Modified Rankin Scale (mRS) score, the change in clinical symptoms, collateral grade, and CTP parameters pre- and postoperatively were compared. RESULTS: The overall level of the change in clinical symptoms in the STA-MCA group was higher than in the EDAS group (p < 0.05). In the operative area, the relative cerebral blood flow (rCBF) was significantly higher whereas the relative time to peak (rTTP) and the relative mean transit time (rMTT) were significantly lower in the STA-MCA and EDAS groups postoperatively than preoperatively (all p < 0.05). In the ipsilateral frontal lobe and basal ganglia, the postoperative rCBF was significantly higher, and the rTTP was significantly lower than the preoperative in the STA-MCA group (all p < 0.05). The postoperative rCBF improvement was higher in each brain area for STA-MCA than in the EDAS group (all p < 0.05). CONCLUSION: Highlighting the utility of CTP, this study demonstrates its effectiveness in assessing postoperative cerebral hemodynamic changes in adult MMD patients. STA-MCA yielded a larger postoperative perfusion area and greater improvement compared to EDAS, suggesting CTP's potential to elucidate symptom variation between two surgical revascularization procedures. CRITICAL RELEVANCE STATEMENT: We analyzed computed tomography perfusion parameters in pre- and postoperative adult Moyamoya disease patients undergoing superficial temporal artery-middle cerebral artery bypass and encephaloduroarteriosynangiosis. Our findings suggest computed tomography perfusion's potential in objectively elucidating symptom variations between these surgical revascularization approaches for MMD. KEY POINTS: • Postoperative perfusion improvement is only confined to the operative area after EDAS. • Besides the operative area, postoperative perfusion in the ipsilateral frontal lobe and basal ganglia was also improved after STA-MCA. • The degree of perfusion improvement in each brain area in the STA-MCA group was generally greater than that in the EDAS group.

Utilization of a lateral flow colloidal gold immunoassay strip based on surface-enhanced Raman spectroscopy for rapid detection of glycinin.

Glycinin is an important storage protein in soybean, but it can also lead to allergic reactions in humans. In this study, based on a low-cost, simple, rapid and portable lateral flow immunoassay test strip, combined with high sensitivity surface enhanced Raman spectroscopy (SERS) technology, a sandwich lateral flow immunochromatographic test strip for rapid detection of soybean allergen glycinin is established. In the experiment, colloidal gold was covalently conjugated with rabbit-derived polyclonal antibodies of glycinin and Raman probe molecule 4-aminothiophenol(4-PATP) to prepare the immunoprobe. The respective optimal PATP and optimal antibody labeling amounts of colloidal gold solution were 1.05 × 10-2mol/L and 4.6 × 10-8mol/L. The detection limit of the test strip for glycinin was 4.87 ng/mL. The recovery rate ranged from 91 to 107 % and the CV was between 3 and 10 %. The test strip underwent no cross-reaction with ß-conglycinin, sesame protein, peanut protein, wheat protein or whey protein. The results of the experiment showed that this method exhibits high sensitivity and specificity.

Mechanism of Yishen Tonglong Decoction inhibiting TLR4/p38 MAPK/NF-κB signaling pathway against prostate cancer via upregulating miR-145-5p / 数字中医药（英文）

@#【Objective】 To investigate the mechanism of Yishen Tonglong Decoction (益肾通癃汤, YSTLD) inhibiting the toll-like receptor 4/p38 mitogen activated protein kinases/nuclear factor kappa-B (TLR4/p38 MAPK/NF-κB) signaling pathway against prostate cancer by up-regulating miR-145-5p. 【Methods】 miRNA microarray technology was used to detect the changes of miRNA expression profile in prostate cancer PC-3 cells treated with YSTLD, and miRNAs with marked differences in miRNA microarray results were screened and validated by real-time polymerase chain reaction (qRT-PCR). Lentiviral transfection of miR-145-5p into prostate cancer PC-3 cells, Cell Counting Kit-8 (CCK8) assay, and scratch assay were adopted to detect the effects of miR-145-5p on prostate cancer PC-3 cell proliferation and migration. qRT-PCR and Western blot were employed to detect the effects of miR-145-5p on TLR4/p38 MAPK/NF-κB signaling pathway and the expression levels of apoptosis-related genes caspase3, tumor necrosis factor-α (TNF-α), Bax, and Bcl-2. qRT-PCR and Western blot were used to detect the effects of serum containing YSTLD on miR-145-5p, TLR4/p38 MAPK/NF-κB signaling pathway, and the expression levels of apoptosis-related genes caspase3, TNF-α, Bax, and Bcl-2. 【Results】 The expression levels of 35 miRNAs in prostate cancer PC-3 cells treated with YSTLD were significantly different from those in the control group, with miR-145-5p being the most significantly different; qRT-PCR validation revealed that the miR-145-5p levels in prostate cancer PC-3 cells treated with YSTLD were significantly higher than those in the DMSO control group (P < 0.05). After lentiviral transfection of miR-145-5p into prostate cancer PC-3 cells, miR-145-5p was found to inhibit the proliferation and migration of prostate cancer PC-3 cells. Overexpression of miR-145-5p up-regulated expression levels of caspase3, TNF-α, and Bax mRNA, and down-regulated expression levels of p38 MAPK, p65 NF-κB, and Bcl-2 mRNA in prostate cancer PC-3 cells (P < 0.05), while up-regulated caspase3 protein expression levels in prostate cancer PC-3 cells and down-regulated expression levels of TLR4, p38 MAPK, and p65 NF-κB protein (P < 0.05). Serum containing YSTLD could up-regulate the expression levels of caspase3, TNF-α, and Bax mRNA, and down-regulate the mRNA expression levels of p38 MAPK, p65 NF-κB, Bcl-2, and TNF receptor-associated factor 1 (TRAF1) in prostate cancer PC-3 cells after intervening prostate cancer PC-3 cells (P < 0.05). Simultaneously, it up-regulated the expression levels of caspase3 protein and down-regulated the protein expression levels of TLR4, p38 MARK, p65 NF-κB, and TRAF1 in prostate cancer PC-3 cells (P < 0.05). 【Conclusion】 YSTLD can promote apoptosis of prostate cancer PC-3 cells by up-regulating the expression level of miR-145-5p and inhibiting TLR4/p38 MAPK/NF-κB signaling pathway, which may be an important mechanism of YSTLD against prostate cancer.

Sleep deprivation leads to further impairment of hippocampal synaptic plasticity by suppressing melatonin secretion in the pineal gland of chronically unpredictable stress rats.

There has been ample research showing that insomnia is a potential trigger of depression as well as a symptom of depression. These two factors contribute to behavioural problems and are closely related to the plasticity of hippocampal synapses. Although depression and insomnia impair hippocampal synaptic plasticity, the mechanism by which this happens remains a mystery. This study aimed to investigate the pathogenesis of insomnia comorbidity in depression and the regulatory effect of venlafaxine combined with melatonin on hippocampal synaptic plasticity in chronic unpredictable mild stress (CUMS) with sleep deprivation (SD) rats. Thus, rats were subjected to 14 days of chronic mild unpredictable stress, gradually acclimated to sleep deprivation on days 12-14. Followed by 21 consecutive days of sleep deprivation, 18 h per day, with daily gavage of venlafaxine (13.5 mg/kg) + melatonin (72 mg/kg) on days 15-36. Venlafaxine + melatonin treatment improves depression-like behaviour, pentobarbital sodium experimental sleep latency, and sleep duration in CUMS +SD rats. In addition to improving depressive-like behaviors, sleep deprivation also upregulates the expression of caspase-specific cysteine protein 3 (Caspase 3) in the pineal glial cells of chronic mild rats, as well as in hippocampal microglia. Expression of ionic calcium-binding adaptor 1 (iba-1), downregulates the secretion of several synaptic plasticity-related proteins, notably cAMP response element binding protein (CREB), glial cell line-derived neurotrophic factor (GDNF), and the synaptic scaffolding protein Spinophiline (Spinophiline). Hematoxylin-eosin staining showed that the structure of the pineal gland and hippocampus was damaged, and Golgi staining showed that the dendrites and spines in the DG area of the hippocampus were destroyed, vaguely aggregated or even disappeared, and the connection network could not be established. Western blot analysis further revealed a positive correlation between low melatonin levels and reduced Spinophiline protein. Interestingly, venlafaxine + melatonin reversed these events by promoting hippocampal synaptic plasticity by regulating melatonin secretion from the pineal gland. Therefore, it exerted an antidepressant effect in sleep deprivation combined with CUMS model rats. Overall, the results of this study suggest that the pathophysiology of depressive insomnia comorbidity is mediated by impaired pineal melatonin secretion and impaired hippocampal synaptic plasticity. In addition, these responses are associated with melatonin secretion from the pineal gland.

[Yishen Tonglong Decoction inhibits the epithelial-mesenchymal transition and Ras/ERK signaling pathway in human prostate cancer DU-145 cells].

Objective: To observe the effect of Yishen Tonglong Decoction (YTD) on the epithelial-mesenchymal transition (EMT) and Ras/ERK signaling pathway in human PCa DU-145 cells and explore its action mechanism. METHODS: We treated human PCa DU-145 cells with normal plasma (the blank control) or plasma containing 5% (low-dose), 10% (medium-dose) and 15% (high-dose) YTD. After intervention, we examined the proliferation of the DU-145 cells in different groups with CCK-8 and their apoptosis by Annexin V/PI double staining. We detected the cell cycle by PI assay, the invasion and migration of the cells using the Transwell chamber and scratch test, and the expressions of the proteins and genes related to the EMT and Ras/ERK signaling pathways in the cells by Western blot and RT-PCR. RESULTS: Compared with the blank control group, high-, medium- and low-dose YTD significantly inhibited the proliferation of the PCa DU-145 cells, decreased their adherence and growth (P < 0.05, P < 0.01), promoted their apoptosis (P < 0.01), regulated their cell cycles (P < 0.05, P < 0.01), and reduced their in vitro invasion and migration abilities (P < 0.05), all in a dose-dependent manner. The results of Western blot and RT-PCR revealed down-regulated protein and mRNA expressions of N-cadherin, zinc finger transcription factor (Snail), Ras, p-ERK1/2 and ERK1/2, but up-regulated protein and mRNA expressions of E-cadherin in the PCa DU-145 cells treated with YTD (P < 0.05, P < 0.01). CONCLUSIONS: Yishen Tonglong Decoction can effectively inhibit the proliferation, promote the apoptosis, regulate the cell cycle and suppress the invasion and migration abilities and EMT process of human PCa DU-145 cells. The mechanism of Yishen Tonglong Decoction acting on PCa may be associated with its inhibitory effect on the EMT process and expression of the Ras/ERK signaling pathway in PCa cells./.

Encephaloduroarteriosynangiosis (EDAS) treatment of moyamoya syndrome: evaluation by computed tomography perfusion imaging.

OBJECTIVE: To explore the value of computed tomography perfusion (CTP) imaging for evaluating the efficacy of encephaloduroarteriosynangiosis (EDAS) treatment of moyamoya syndrome (MMS). METHODS: Forty-three patients with MMS (48 hemispheres) who received EDAS treatment were examined using CTP and DSA before and after surgery. CTP of the ipsilateral cortex, contralateral mirror area, and pons region were measured, and the relative cerebral blood flow (rCBF) and volume (rCBV), mean transit time (rMTT), and time-to-peak (rTTP) were calculated. Based on postoperative DSA, 48 hemispheres were apportioned to two groups based on rich (grades 2, 3) or poor (grades 0, 1) collateral vessel formation, and the pre- and post-operative differences in perfusion changes were compared. The association between clinical outcome, CTP, and the degree of DSA collateral vessels was explored. RESULTS: rCBF and rMTT significantly improved in both the poor and rich collateral vessel formation groups (n = 21 and 27, respectively), while rTTP significantly improved only in the latter. Postoperative CTP improved in the rich and the grade 1 collateral vessel groups (p < 0.01). The clinical improvement was consistent with the improvement of CTP (p = 0.07), but less consistent with the degree of collateral angiogenesis (p = 0.003). CONCLUSION: CTP can quantitatively evaluate the improvement of brain tissue perfusion in the operated area after EDAS. Brain tissue perfusion in operated areas improved regardless of postoperative rich or poor collateral vessel formation observed via DSA. A significant improvement in rTTP in the operated area may indicate the formation of abundant collateral vessels. KEY POINTS: • CTP showed that brain tissue perfusion in the operated area after EDAS improved regardless of rich or poor collateral vessel formation observed via DSA. • Significant improvement of rTTP in the operated area may indicate the formation of abundant collateral vessels.

Computed tomographic angiography may be used for assessing the dilatation of the anterior choroidal and posterior communicating arteries in patients with moyamoya syndrome.

OBJECTIVE: To evaluate the feasibility of CT angiography (CTA) for assessing anterior choroidal artery (AChA) and posterior communicating artery (PComA) dilatation in patients with moyamoya syndrome (MMS). METHODS: Eighty-eight MMS patients who underwent digital subtraction angiography (DSA) and CTA within 1 month were enrolled. The AChA was graded using both DSA and CTA. Given the features of dual blood supply, DSA was firstly used for grading of the PComA. Then, the calibers of PComA, P1 or P2 segment of the posterior cerebral artery (PCA), were recorded from CTA. Taking DSA as a reference standard, the optimal cutoff values of the PComA/P1 or PComA/P2 were calculated to determine the dilatation of PComA. Both the AChA and PComA were classified as extreme dilatation (ED, grade 2) or non-extreme dilatation (NED, grade 0 or 1). RESULTS: The AChA was evaluated in 149 affected hemispheres of 88 patients while the PComA was evaluated in 70 affected hemispheres of 49 patients. The sensitivity and specificity of CTA in diagnosing AChA-ED were 92% and 93.5% respectively. Both the PComA/P1 (p < 0.001) and PComA/P2 (p = 0.4) ratios were increased in the PComA-ED group with the former yielding a better detecting performance than the latter (AUC = 0.92 vs 0.85, p = 0.046). When using 0.71 as a cutoff value, the sensitivity and specificity of the PComA/P1 ratio for diagnosis of PComA-ED cases were 91.3% and 83.3% respectively. CONCLUSIONS: CTA could be used for the AChA classification in MMS patients, while a PComA/P1 ratio greater than 0.71 indicates the existence of PComA-ED. KEY POINTS: • CTA showed a high sensitivity, specificity, and accuracy in diagnosing AChA-ED in patients with MMS. • PComA/P1 ratio greater than 0.71 on CTA signified an extremely dilated PComA. • CTA could be used to assess the dilatation of AChA and PComA in MMS patients, especially for routine postoperative follow-up.

Pingyang Jiangya Fang pretreatment reduces the blood pressure of spontaneously hypertensive rats with Liver-Yang hyperactivity syndrome via ROS/Akt oxidative stress pathway.

BACKGROUND: ROS/Akt pathway oxidative stress refers to a procedural response that activates various stress-sensitive signaling pathways in the cell thereby inducing insulin resistance (IR), and is closely related to high blood pressure. This study aims to investigate the effects of Pingyang Jiangya Fang (PYJYF) on the ROS/Akt pathway oxidative stress response in spontaneously hypertensive rats (SHRs) with Liver-Yang hyperactivity syndrome. METHODS: Except for the Wistar-Kyoto (WKY) and SHR groups, the other groups' rats were administered with Fuzi Decoction, to duplicated the model of hyperactivity of Liver-Yang. These rat's scleral color of deepens and become red, and rat's degree of irritability reached to above II degree. Compared with control group, rat's water consumption increased significantly, rotation tolerance time reduced significantly, pain threshold reduced significantly, rat tail vein systolic blood pressure (SBP) increased significantly. It shows that the model rats with hyperactivity of Liver-Yang have successfully replicated. Then, the corresponding drugs in the positive medicine group (candesartan cilexetil tablets) and the high, middle, and low dose groups of PYJYF (PYJYF-H, PYJYF-M, PYJYF-L) were administered. This work detected and analyzed behavior, SBP, renin-angiotensin system (RASS), expression of skeletal muscle angiotensin II type 1 receptor/angiotensin II type 2 receptor (AT1R/AT2R), IR index, renal/renal vascular/skeletal muscle tissue damage in each group. Moreover, we identified the target effect of point composition in the ROS/Akt signaling pathway. RESULTS: Unlike the WKY and SHR groups, the model group exhibited the phenotype of Liver-Yang hyperactivity syndrome, markedly increased SBP, hyper-RASS system, increased urine N-acetyl-ß-Dglucosidase (NAG) and microalbuminuria (m-ALB), increased AT1R/AT2R ratio of skeletal muscle, IR, concomitant renal and skeletal muscle damage, as well as significant upregulation in the expression of ROS/ Akt signaling factor. PYJYF-H and PYJYF-M improved Liver-Yang hyperactivity syndrome, reduced SBP, the RASS system, urine NAG and m-ALB, AT1R/AT2R ratio, IR index, repair renal, renal vascular, and skeletal muscle tissue injury, as well as downregulated the expression of ROS/Akt signal factor. CONCLUSIONS: PYJYF reduces blood pressure by inhibiting ROS/Akt pathway oxidative stress and reducing IR.

LncRNA KCNQ1OT1 sponges miR-15a to promote immune evasion and malignant progression of prostate cancer via up-regulating PD-L1.

BACKGROUND: We focused on the KCNQ1OT1/miR-15a/PD-L1 axis and explored its significance in regulating immune evasion and malignant behaviors of prostate cancer (PC) cells. METHODS: The expression levels of KCNQ1OT1, miR-15a, PD-L1, and CD8 in cells or tissues were examined by RT-qPCR, western blot or immunohistochemistry (IHC) assays. The direct regulations between KCNQ1OT1, miR-15a and PD-L1 were validated by luciferase reporter assay. PC cells were co-cultured with CD8+ T cells to study the immune evasion. Proliferation, apoptosis, migration and invasion abilities were detected by MTT, flow cytometry, wound healing and Transwell assays, respectively. The cytotoxicity of CD8+ T cells was determined by LDH cytotoxicity Kit. Epithelial-mesenchymal transition (EMT) and Ras/ERK signaling markers were evaluated by western blot. RESULTS: KCNQ1OT1, PD-L1 and CD8 were increased, while miR-15a was decreased in PC tissues. MiR-15a directly bound to the 3'-UTR of PD-L1 and inhibited the expression of PD-L1. Overexpressing miR-15a in PC cells was sufficient to promote cytotoxicity and proliferation, while inhibit apoptosis of CD8+ T cells, and also suppressed viability, migration, invasion and EMT while promoted apoptosis of PC cells. The above anti-tumor effects of miR-15a were reversed by overexpressing PD-L1. KCNQ1OT1 sponged miR-15a and released its inhibition on PD-L1. Functionally, KCNQ1OT1 in PC cells was essential for suppressing the cytotoxicity of CD8+ T cells and maintaining multiple malignant phenotypes of PC cells. The Ras/ERK signaling was suppressed after overexpressing miR-15a or knocking down KCNQ1OT1. CONCLUSIONS: LncRNA KCNQ1OT1 sponges miR-15a to promote immune evasion and malignant progression of PC via up-regulating PD-L1.

Evaluation of Hemodynamics Before and After Revascularization in Hemorrhagic Moyamoya Disease: A Computed Tomography Perfusion Imaging Case Study.

OBJECTIVE: To explore the feasibility of computed tomography perfusion imaging (CTP) for evaluating hemodynamics in hemorrhagic moyamoya disease (MMD). METHODS: The retrospective analysis included 25 patients with hemorrhagic MMD who underwent brain CTP examination. Two experienced radiologists manually delineated regions of interest (ROIs) in the bilateral frontal lobe, temporal lobe, brain tissue adjacent to the hemorrhagic foci, and brainstem as a control region. The perfusion values for all ROIs were extracted, including cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT), and time to peak (TTP). Subsequently, the differences in perfusion values for different brain tissues were compared between the hemorrhagic side and the nonhemorrhagic side. For patients who underwent revascularization surgery, differences in perfusion values from before to after surgery were determined in brain tissues on the ipsilateral side. RESULTS: CBF in the area around the hematoma and the lateral temporal lobe on the hemorrhage hemisphere was lower than that on the contralateral side, whereas TTP and MTT were higher. Among the 14 patients who underwent revascularization, CBF and CBV in the postoperative temporal lobes were higher than the preoperative values, whereas TTP and MTT were lower. CBF and CBV in the frontal lobe were higher after the operation. CONCLUSIONS: Hemorrhagic MMD results in cerebral ischemia, and CTP could be used to localize such ischemic brain tissue and objectively evaluate the changes in cerebral hemodynamics with revascularization.

Significance of nuclear magnetic resonance combined with Ki-67 and VEGF detection in the diagnosis and prognosis evaluation of brain glioma.

PURPOSE: To investigate the significance of nuclear magnetic resonance (NMR) combined with Ki-67 and vascular endothelial growth factor (VEGF) detection in the diagnosis and prognosis evaluation of brain glioma. METHODS: 78 patients with brain glioma treated at the Affiliated Hospital of Jining Medical University from January 2015 to August 2017 were studied. All patients underwent the NMR diffusion tensor imaging examination. The expressions of Ki-67 and VEGF in brain glioma tissues were detected using immunohistochemistry and survival analyses were performed for patients in high apparent diffusion coefficient (ADC) group and low ADC group, high-expression Ki-67 and VEGF group and low-expression Ki-67 and VEGF group before treatment. Moreover, the value of combined diagnosis in the prognosis evaluation of patients was analyzed. RESULTS: NMR diffusion tensor imaging showed that the fractional anisotropy (FA) value in the tumor enhancement region on the affected side was significantly lower than in the contralateral normal region, but ADC was significantly higher in the contralateral normal region; the FA value in grade I-II brain glioma enhancement region was higher compared with grade III-IV glioma enhancement region, but ADC was lower in the grade-III-IV glioma enhancement region (p<0.05). The low expression rates of Ki-67 and VEGF in patients with grade I-II brain glioma were significantly higher than in patients with grade III-IV glioma (p<0.05). After treatment, the 3-year survival rate of high ADC group was lower than that of low ADC group, and the 3-year survival rate of high-expression Ki-67 and VEGF group was also obviously lower than that in low-expression Ki-67 and VEGF group (p<0.05). Besides, the area under the receiver operating characteristic (ROC) curve of NMR combined with Ki-67 and VEGF detection in the prediction of patient prognosis was 0.906, the sensitivity 91.6%, and the specificity 89.5%. CONCLUSION: NMR diffusion tensor imaging has a high application value in the diagnosis of brain glioma. The expressions of Ki-67 and VEGF are related to the pathological grade of glioma, which can be used as biological indexes for the diagnosis of glioma. Moreover, the combined detection of the three items can not only accurately determine the grade of glioma malignancy, but also effectively evaluate the prognosis of patients, thus providing a scientific basis for the selection of therapeutic regimen.

Intracranial Arterial Fenestration and Risk of Aneurysm: A Systematic Review and Meta-Analysis.

BACKGROUND: Previous studies have been inconsistent regarding risk for intracranial aneurysm related to intracranial arterial fenestration. We conducted a meta-analysis to examine the association between intracranial arterial fenestration and risk of aneurysm. METHODS: We performed a systematic review of PubMed and Embase through August 2017 for potentially relevant articles. Summary odds ratios with 95% confidence intervals were pooled using a random-effects model. RESULTS: Of 446 articles found, 7 were selected for meta-analysis. Pooled odds ratios revealed an increased risk of aneurysm owing to fenestration of 1.50 (95% confidence interval, 0.61-3.71; P = 0.38). Subgroup analyses based on the population presenting with various indications suggested that pooled odds ratios indicated a significant increase in risk for aneurysm of 2.43 (95% confidence interval, 1.04-5.69; P = 0.04). CONCLUSIONS: Our findings indicate that intracranial arterial fenestration may be associated with increased risk for aneurysm formation.

Application of CT perfusion to assess hemodynamics in symptomatic Moyamoya syndrome: focus on affected side and parameter characteristic.

OBJECTIVE: Vascular and hemodynamic changes were not consistent in symptomatic and non-symptomatic cerebral hemisphere in patients with symptomatic moyamoya syndrome (MMS). Thus, the purpose of this study is to evaluate the hemodynamic difference between symptomatic and non-symptomatic cerebral hemisphere in patients with symptomatic MMS. METHODS: Patients who were diagnosed with symptomatic MMS were retrospectively collected. All cases underwent CTP examination. Regions of interest (ROIs) were chosen in the mirroring bilateral frontal lobes, temporal lobes, the basal ganglia, and the brainstem as control region. The relative perfusion parameter values of symptomatic side were compared with non-symptomatic side. RESULTS: Of the 40 patients, 33 patients were taken into assessment. In all cases (n = 33), rCBF, rMTT, and rTTP in all regions of interest (ROIs) of the symptomatic side were significantly different from those of contralateral side. In unilateral MMS patients (n = 7), rCBF values were not significantly different between two sides in the temporal lobe and basal ganglia area; rTTP values were significantly higher in the symptomatic side. rMTT values were significantly higher only in the temporal lobe of symptomatic side. In bilateral MMS patients (n = 26), rCBF and rMTT in all ROIs of the symptomatic side were significantly different from those of contralateral side. However, there were no significant differences between two sides in all ROIs on rTTP values. CONCLUSIONS: This study demonstrates that rCBF and rMTT were more sensitive than rTTP for evaluating hemodynamic changes in patients with symptomatic bilateral MMS. Furthermore, patients with unilateral MMS may have a preserved rCBF compared to those with bilateral disease.

De Novo Mutation of Paternal IGF2 Gene Causing Silver-Russell Syndrome in a Sporadic Patient.

Silver-Russell syndrome (SRS) is a rare, but well-recognized disease characterized by growth disorder. To date, there are two reports arguing IGF2 mutation for the onset of SRS. Herein, we present another sporadic case harboring IGF2 mutation. The male proband was the first and only child of a non-consanguineous Chinese couple. He was small for gestational age, with relative macrocephaly at birth. Severe feeding difficulties, low feeding, and growth retardation were revealed during neonatal period. At 4.5 years old, obvious body asymmetry was noted. Whole exome sequencing identified a novel de novo c.101G > A (p.Gly34Asp, NM_000612) variant in IGF2 and Sanger sequencing validated the variant. Amplification refractory mutation system polymerase chain reaction demonstrated that the IGF2 variant was on the paternal allele. Alignment shows the variant is evolutionarily conserved. Structural modeling argues that the variant site might be important for the binding of IGF2 to its receptor. Our study provides further evidence that IGF2 mutation may be another mechanism of SRS, and we consider that IGF2 should be included in a disease specific gene panel in case it is designed for SRS routine diagnostics.

Functional expression of the globular domain of human adiponectin in Pichia pastoris.

Adiponectin is an adipokine that predominantly synthesized and secreted from adipocytes mainly in the white adipose tissue. Here, we report that we have successfully expressed human gAdiponectin (the globular domain of adiponectin) in the methylotrophic yeast Pichia pastoris after codon optimization and established the purification procedure. The human gAdiponectin gene was designed and synthesized by PCR according to the P. pastoris preferred codons, and then inserted into the P. pastoris pPIC9K expression vector. The plasmid was electroporated into the P. pastoris strain GS115 and only the G418 resistance colonies could produce the gAdiponectin. After fermentation and purification, we could get 1.2g of recombinant gAdiponectin (purity is approximately 95%) from a 24 L culture media. The recombinant gAdiponectin is fully functional as evidenced by induction the phosphorylation of ACC in differentiated C2C12 myotubes, significantly lowering the blood glucose level and accelerating the clearance of free fatty acid in animal models.

Lactacystin inhibits 3T3-L1 adipocyte differentiation through induction of CHOP-10 expression.

Hormonal induction triggers a cascade leading to the expression of CCAAT/enhancer-binding protein(C/EBP)alpha and peroxisome proliferator-activated receptor (PPAR) gamma, C/EBPalpha, and PPARgamma turns on series of adipocyte genes that give rise to the adipocyte phenotype. Previous findings indicate that C/EBPbeta, a transcriptional activator of the C/EBPalpha and PPARgamma genes, is rapidly expressed after induction, but lacks DNA-binding activity and therefore cannot activate transcription of the C/EBPalpha and PPARgamma genes early in the differentiation program. Acquisition of DNA-binding activity of C/EBPbeta occurs when CHOP-10, a dominant-negative form of C/EBP family members, is down-regulated and becomes hyperphosphorylated as preadipocytes traverse the G1-S checkpoint of mitotic clonal expansion. Evidences are presented in this report that lactacystin, a proteasome inhibitor, up-regulated the CHOP-10 expression, blocked the DNA-binding activity of C/EBPbeta, and subsequently inhibited MCE as well as adipocyte differentiation.

Identification of a peroxisome proliferator responsive element (PPRE)-like cis-element in mouse plasminogen activator inhibitor-1 gene promoter.

PAI-1 is expressed and secreted by adipose tissue which may mediate the pathogenesis of obesity-associated cardiovascular complications. Evidence is presented in this report that PAI-1 is not expressed by preadipocyte, but significantly induced during 3T3-L1 adipocyte differentiation and the PAI-1 expression correlates with the induction of peroxisome proliferator-activated receptor gamma (PPARgamma). A peroxisome proliferator responsive element (PPRE)-like cis-element (-206TCCCCCATGCCCT-194) is identified in the mouse PAI-1 gene promoter by electrophoretic mobility shift assay (EMSA) combined with transient transfection experiments; the PPRE-like cis-element forms a specific DNA-protein complex only with adipocyte nuclear extracts, not with preadipocyte nuclear extracts; the DNA-protein complex can be totally competed away by non-labeled consensus PPRE, and can be supershifted with PPARgamma antibody. Mutation of this PPRE-like cis-element can abolish the transactivation of mouse PAI-1 promoter mediated by PPARgamma. Specific PPARgamma ligand Pioglitazone can significantly induce the PAI-1 expression, and stimulate the secretion of PAI-1 into medium.