Changes of photopic negative response of multifocal electroretinogram after treatment with Aflibercept in patients with diabetic macular edema / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To observe the changes of photopic negative response(PhNR)of multifocal electroretinogram(mf-ERG)in patients with diabetic macular edema(DME)before and after treatment with Aflibercept.METHODS: A total of 37 patients(37 eyes)with DME who visited the Aier Eye Hospital of Wuhan University(Wuhan Aier Eye Hospital)from May 2019 to June 2022, and 0.05 mL of aflibercept was injected per month for consecutive 3mo were included in this retrospective cohort study. Another 20 cases(20 eyes)with normal physical examination to exclude related eye diseases were selected as the control group. The PhNR amplitude of mf-ERG, best-corrected visual acuity(BCVA; LogMAR), central retinal thickness(CRT), capillary plexus in macular area and vessel density(CPVD)of the participants between the two groups were compared before and after treatment.RESULTS: The PhNR amplitude of mf-ERG in DME patients before treatment(201.69±80.92nV)was significantly lower than that in the normal control group(398.87±77.92nV; P<0.01), and the average PhNR amplitude of mf-ERG in DME patients at 6mo after treatment was significantly higher than that before treatment(P=0.036), but it was still significantly lower than the normal control group at 6mo after treatment(P=0.031). In addition, the BCVA(LogMAR)of DME patients increased from 0.64±1.33 to 0.37±1.39(P=0.021)at 6mo after treatment, and CPVD significantly increased compared to that before treatment(P=0.029). Meanwhile, the PhNR amplitude of mf-ERG in DME patients at 6mo after treatment was positively correlated with CPVD at 6mo after treatment(r=0.448, P=0.043), and negatively correlated with BCVA(LogMAR)and CRT(r=-0.647, P=0.011; r=-0.337, P=0.032).CONCLUSION: The PhNR amplitude of mf-ERG in DME patients increased significantly after receiving aflibercept, and it can be used to observe and evaluate the functional changes of retinal ganglion cells in DME patients.

[Clinical Study of miRNAs Derived from Serum Exosomes in Multiple Myeloma].

OBJECTIVE: To investigate the expression level and prognostic value of miR-21, miR-18a, miR-146a, and Let-7b derived from serum exosomes in patients with multiple myeloma (MM). METHODS: Serum exosomes were extracted from 57 MM patients and 20 healthy persons using ExoQuick exosome precipitation solution kit, and the relative expression level of miR-21, miR-18a, miR-146a, and Let-7b derived from serum exosomes was measured by RT-qPCR. Correlations of the expression levels of all miRNAs mentioned above with routine laboratory parameters were analyzed by Spearman correlation analysis. The relationship between the expression level of miR-21, miR-18a, miR-146a, and Let-7b derived from serum exosomes and overall survival of patients with MM was analyzed using the Kaplan-Meier survival curve. RESULTS: The expression levels of miR-21, miR-18a, and Let-7b derived from serum exosomes in patients with MM were significantly lower than those in the normal control group (P<0.001), while the expression level of miR-146a between the two groups was not significantly different (P>0.05). The expression level of miR-21 was strongly negatively correlated with serum ß2-microglobulin concentration (r=-0.830), and weakly negatively correlated with serum creatinine, corrected serum calcium, and cystatin C (r=-0.488, -0.282, -0.627). The expression levels of Let-7b and miR-18a were also weakly negatively correlated with the corrected serum calcium, ß2-microglobulin, and cystatin C concentration (r=-0.305, -0.362, -0.461; -0.317, -0.542, -0.434). However, there was no significant correlation between the expression level of miR-146a and routine laboratory parameters in MM patients. The overall survival rate of MM patients with low expression level of miR-21, miR-18a, and Let-7b significantly decreased compared with high expression level group (P<0.05), however, the expression level of miR-146a was not related to the overall survival rate. CONCLUSION: Aberrant low expression levels of miR-21, miR-18a, and Let-7b derived from serum exosomes exist in patients with MM, which are associated with a worse overall survival rate.

The mechanism of RCE-4, an active ingredient of Reineckia carnea, in combination with celecoxib on the anti-proliferation of cervical cancer Ca Ski cells / 药学学报

This research explored the synergistic effects and the potential mechanisms of RCE-4 and various nonsteroidal anti-inflammatory drugs (NSAIDs) on the proliferation of cervical cancer Ca Ski cells. The MTT assay and CalcuSyn V2.0 software were used to detect cell proliferation and calculate the combination index (CI); the expression levels of various proteins were analyzed using Western blot assay; mitochondrial membrane potential (MMP) was assessed using JC-1 staining; acridine orange/ethidium bromide (AO/EB) double-fluorescence staining was used to detect the apoptosis of Ca Ski cells; a co-immunoprecipitation (Co-IP) assay was used to analyze the relative content of Bcl-2-Beclin 1 complex in Ca Ski cells. The results demonstrate that the combination of RCE-4 and NSAIDs increases the inhibition of Ca Ski cells compared to the single-RCE-4 group, and celecoxib provided the best synergistic effect among the four NSAIDs tested, with a CI of 0.32. The combination of RCE-4 and celecoxib significantly down-regulated the expression of cyclooxygenase-2 (COX-2) and nuclear transcription factor-κB (NF-κB), and promoted the expression of non-steroidal anti-inflammatory drugs activity gene-1 (NAG-1). In addition, autophagy induced by RCE-4 was markedly inhibited in combination with celecoxib, which was associated with down-regulation of the expression of microtubule-associated protein 3 (LC3)-II, Beclin 1, p62 and autophagy-related gene (ATG) 3/4B/5/7/14. RCE-4-induced apoptosis was significantly enhanced by altering the depolarization of mitochondrial membrane potential and the expression of B cell lymphoma-2 (Bcl-2), B cell lymphoma-xl (Bcl-xl), Bcl-2 associated X protein (Bax), Bcl-2/Bcl-xl-associated death promoter (Bad) and cleaved cysteinyl aspartate specific proteinase (cleaved-caspase) 3/7/9. Furthermore, the formation of the Bcl-2-Beclin 1 complex was significantly inhibited in Ca Ski cells treated with RCE-4 in combination with celecoxib. Taken together, this research shows that the combination of RCE-4 and celecoxib has a significant synergistic effect on the proliferation of Ca Ski cells by promoting apoptosis, inhibiting autophagy and disturbing the formation of the Bcl-2-Beclin 1 complex, which may be a novel strategy to increase the sensitivity of anti-cervical cancer drugs.

Effects of RCE-4, an active component of Reineckia carnea, on regulation of cell cycle arrest in human cervical cancer Ca Ski cells by triggering endoplasmic reticulum stress / 中国药理学通报

Aim: To explore the mechanism of RCE-4, an active constituent isolated from Reineckia carnea on anti-proliferation activity of human cervical neoplasm Ca Ski cells. Methods Etramethylazolyl blue method (MTT) and clone formation assay were adopted to observe the inhibitory effects of RCE-4 on the growth of Ca Ski cells, and its half maximal inhibitory concentration (IC

Association between CD137 and ischemia-reperfusion injury in patients with acute ST-segment elevation myocardial infarction / 中华心血管病杂志

Objective: To explore the relationship between the levels of serum soluble CD137 (sCD137) and membrane-bound CD137 (mCD137) and the occurrence of ischemia reperfusion injury (IRI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods: This is a cross-sectional study. Consecutive patients with acute STEMI, who underwent emergency percutaneous coronary intervention (PCI) in the Department of Cardiology, Jiangsu University Affiliated Hospital from May 2019 to September 2020, were enrolled. According to the absence or presence of IRI, patients were divided into IRI group and non-IRI group. Clinical data of the two groups were collected and compared. sCD137 level was detected by enzyme linked immunosorbent assay. Ficoll density gradient centrifugation was used to separate peripheral blood mononuclear cells (PBMC) and RNA was extracted, mCD137 mRNA expression level was detected by PCR. Serum sCD137 levels and the mCD137 mRNA levels of PBMC before, after PCI and 24 hours after PCI were compared. The correlation between serum sCD137 level, PBMC mCD137 mRNA level and clinical indicators was observed. The univariate and multivariate logistic binary regression analyses were performed to evaluate the related risk factors of IRI. ROC curve was used to analyze the predictive value of defined parameters for IRI. Results: A total of 112 STEMI patients were enrolled. There were 42 cases (of which 33 were males (78.6%), mean age was (58.6±12.7) years) in non-IRI group and 70 cases(of which 56 were males (80.0%), mean age was (64.5±11.6) years) in IRI group. Compared with the non-IRI group, patients in the IRI group had longer hospital stays, older age, lower rates of obesity, lower systolic and diastolic blood pressure at admission, higher proportion of the the right coronary artery as culprit vessel, lower rate of the use of angiotensin-converting enzyme inhibitor/angiotensin-Ⅱ receptor blocker/angiotensin receptor neprilysin inhibitor, higher levels of urea nitrogen and creatinine, lower glomerular filtration rate, lower triglycerides, higher D-dimer and B-type natriuretic peptidemax, higher proportion of Killip grade Ⅳ and cardiovascular adverse events (all P<0.05). sCD137 levels at the preoperative, postoperative and 24 hours after surgery were significantly higher in the IRI group than in the non-IRI group, while the mRNA levels of CD137 was similar between the two groups. The level of sCD137 in patients after PCI was lower than that before operation, the level of mCD137 mRNA was higher than that before operation (P<0.05). Serum sCD137 levels were positively correlated with hospitalization days, age, B-type natriuretic peptide, creatinine, ischemic time, C Reactive protein (CRP) and CRP/albumin (P<0.05), and negatively correlated with body mass index, glomerular filtration rate and albumin (P<0.05). The mCD137 mRNA expression level of PBMC was positively correlated with hospital stay, age, B-type natriuretic peptide, ischemic time, CRP and CRP/albumin (P<0.05), and negatively correlated with body mass index, glomerular filtration rate, albumin (P<0.05). Multivariate logistic regression analysis showed that higher sCD137 (OR=1.038, 95%CI: 1.009-1.069), aspartate aminotransferase, (OR=1.029, 95%CI: 1.009-1.050) and lower albumin (OR=0.829, 95%CI: 0.703-0.829) before surgery were independent risk factors of IRI (P<0.05). Receiver operating characteristic curve showed that the area under the curve of sCD137 was 0.672 (95%CI: 0.574-0.770, P=0.002) for the prediction of IRI, the best cut-off value was 28.43×10-3 μg/L with sensitivity of 95.2% and specificity of 48.6%. Conclusion: The significantly increased level of sCD137 in acute STEMI patients is positively correlated with reperfusion injury, which is an independent risk factor of IRI and may be related to the prognosis of patients with IRI.

Role of SIRT1/Nrf2 signaling pathway in sleep deprivation-induced cognitive dysfunction in rats / 中华麻醉学杂志

Objective:To evaluate the role of silent information regulator 1 (SIRT1)/nuclear factor E2 related factor 2 (Nrf2) signaling pathway in sleep deprivation-induced cognitive dysfunction in rats.Methods:Thirty-six adult male Sprague-Dawley rats, aged 54-56 weeks, weighing 600-750 g, were divided into 3 groups ( n=12 each) using a random number table method: control group (group C), sleep deprivation group (group SD) and sleep deprivation+ SIRT1 agonist Srt1720 group (group SD+ Srt). Sleep deprivation model was established by modified multi-platform water environment method.Srt1720 10 mg/kg was injected intraperitoneally every 12 h starting from 24 h before establishing the model in group SD+ Srt, while the equal volume of 0.9% normal saline was injected intraperitoneally in C and SD groups.After the end of sleep deprivation, Morris water maze test was performed to evaluate the cognitive function.Animals were then sacrificed, and their hippocampi were removed for determination of neuronal degeneration rate in hippocampal CA1 region (using HE staining), the apoptosis rate in hippocampal CA1 (using TUNEL assay ), the expression of SIRT1 and Nrf2 in hippocampal CA1 (by immunohistochemistry) and the contents of reactive oxygen species (ROS) and superoxide dismutase (SOD) (by microplate method). Results:Compared with group C, the escape latency was significantly prolonged, the time of staying at the platform quadrant was shortened, and the frequency of crossing the platform was decreased on 2-4 days, the apoptotic rate and neuronal degeneration rate were increased, the expression of SIRT1 and Nrf2 was down-regulated, ROC content was increased, and SOD content was decreased in SD and SD+ Srt groups ( P<0.05). Compared with group SD, the escape latency was significantly shortened, the time of staying at the platform quadrant was prolonged, and the frequency of crossing the platform was increased on 3 and 4 days, the apoptotic rate and neuronal degeneration rate were decreased, the expression of SIRT1 and Nrf2 was up-regulated, ROC content was decreased, and SOD content was increased in group SD+ Srt ( P<0.05). Conclusions:Sleep deprivation can induce oxidative stress response in hippocampus by inhibiting the activation of SIRT1/Nrf2 signaling pathway, leading to cognitive dysfunction in rats.

Analysis of Bacterial Flora Structure of Patients with Cerebral Hemorrhage Due to Hyperactivity of Liver-Yang by 16S rRNA Gene Sequencing Technique / 中国实验方剂学杂志

Objective: To explore the difference of intestinal flora between the patients with acute cerebral hemorrhage due to hyperactivity of liver-Yang and the healthy population. Method: The fecal samples of 9 patients with acute cerebral hemorrhage due to hyperactivity of liver-Yang from the first affiliated hospital of Guangzhou university of traditional Chinese medicine in 2018 were selected as observation group,and 6 stool samples from healthy subjects were selected as the control group.The total bacterial DNA was extracted from the two groups of samples,amplified according to the 16S rRNA V4 region,and paired-end sequencing was performed on the Illumina MiSeq platform.The sequencing results were analyzed by bioinformatics analysis software.The flora composition and structure of the samples from two groups were compared. Result:Venn analysis of operational taxonomic units(OTU) showed significant difference in OTU numbers between the observation group and control group.Partial least squares-discriminant analysis(PLS-DA) showed that there was a significant difference in the composition of intestinal flora between patients with acute cerebral hemorrhage and healthy subjects.On the analysis of species and abundance,at the classification level of phylum,compared with the control group,the ratio of relative abundance values of Firmicutes and Bacteroidetes(F/B) in the observation group was significantly increased,and the relative abundance of Verrucomicrobia was significantly decreased(PPrevotella,Bacteroides,Akkermansia,Blautia and Acidaminococcus(PPBacteroides and Prevotella(B/P) in the observation group was significantly higher than that of the control group;at the classification level of species,there were significant differences between the two groups in P. copri,A. muciniphila,B. ovatus,B. fragilis and Ruminococcus callidus(PPConclusion:Acute cerebral hemorrhage due to hyperactivity of liver-Yang is associated with structural disorder of intestinal flora,which is closely related to the decrease in relative abundance of P. copri and A. muciniphila.

Content Determination of 6 Flavonoids in Epimedium brevicornu from Shenqi Yanshen Granules Based on HPLC-QAMS / 中国药房

OBJECTIVE： To establish a HPLC method for simultaneous determination of 6 flavonoids in Epimedium brevicornu from Shenqi yanshen granules， such as epimedin A1， epimedin A， epimedin B， epimedin C， icariin and baohuoside Ⅰ. METHODS： HPLC method was adopted. The determination was performed on Waters Symmetry C18 column with mobile phase consisted of acetonitrile-water （gradient elution） at the flow rate of 1.0 mL/min； the column temperature was 25 ℃， and detection wavelength was 270 nm. The sample size was 10 μL. Relative correction factors （fk/s） of each component to icariin （reference substance） were established by multi-point correction method and slope correction method on the basis of external standard method to calculate the contents of each component. The contents of 6 flavonoids in 4 batches of Shenqi yanshen granules determined by HPLC external standard method were compared with by multi-point correction method and slope correction method. Feasibility and accuracy of quantitative analysis of multi-components by single-marker （QAMS） were validated. RESULTS： The linear range of epimedin A1， epimedin A， epimedin B， epimedin C， icariin and baohuoside Ⅰ were 2.03-50.80 μg/mL （r＝0.999 5）， 4.34-108.60 μg/mL （r＝0.999 5）， 2.26-56.40 μg/mL （r＝0.999 5）， 4.14-103.60 μg/mL （r＝0.999 5）， 4.24-106.00 μg/mL （r＝0.999 5）， 1.78-44.60 μg/mL （r＝0.999 5）， respectively， the limits of detection were 65.80， 71.49， 74.26， 68.79， 70.56， 86.09 ng/mL， respectively； the limits of quantification were 196.62， 213.63， 223.72， 208.46， 215.96， 255.88 ng/mL， respectively； RSDs of precision， stability （24 h）， reproducibility tests were less than 2％ （n＝6）， respectively. The average recoveries were 96.03％-99.04％ （RSDs were 0.65％-1.04％， n＝6）. By multi-point correction method， fk/s of epimedin A1， epimedin A， epimedin B， epimedin C and baohuoside Ⅰ were 0.837， 0.818， 0.845， 0.831， 1.387， respectively； by slope correction method， fk/s of them were 0.835， 0.815， 0.851， 0.829， 1.419， respectively. There was no significant difference in content determination results between two correction methods of QAMS and external standard （P＞0.05）. CONCLUSIONS： Established HPLC- QAMS method is accurate and suitable for the quality control of epimedin A1， epimedin A， epimedin B， epimedin C， icariin and baohuoside Ⅰ in E. brevicornu from Shenqi yanshen granules.

Optimization of One-step Pelletization Technology of Rouganbao Granules with Orthogonal Test Methodology / 中国中医药信息杂志

Objective To optimize the process conditions of one-step pelletization of Rouganbao Granules. Methods The factors influencing the pelletization of Rouganbao Granules were investigated by L9(34)orthogonal test, with the indexes of forming rate and fluidity. Results The spraying speed had the greatest effect on one-step pelletization, followed by atomization pressure and material temperature, and wind temperature had the smallest effect. At last, the best process parameters were relative density 1.15 (60 ℃), spray speed 55 mL/min, atomization pressure 0.25 MPa, wind temperature 75 ℃, material temperature 55 ℃, and critical relative humidity was 63%. Conclusion One-step pelletization technology can improve the preparation level and product quality, which can be used for the industrial production of Rouganbao Granules.

Content Determination of 5 Anthraquinones in Huangzhi Neiyi Capsules by QAMS / 中国中医药信息杂志

Objective To determine the contents of 5 anthraquinones (aloe-emodin, phein, emodin, chrysophanol, physcion)in Huangzhi Neiyi Capsule through a quantitative analysis of multi-component by single-marker(QAMS), To determine the application value of QAMS by comparing with external standard method (ESM). Methods HPLC was performed at 30 ℃ with Waters Symmetry C18 column (4.6 mm × 250 mm, 5 μm), methanol-0.4% phosphoric acid (85:15) as mobile phase at the wavelength 440 nm, and flow rate of 1.000 mL/min. The relative correction factors of other anthraquinones were calculated respectively by emodin as the reference. Results The values calculated by QAMS were compared with the values measured by ESM, and no significant difference was showed (RSD<5%). Conclusion For the availability and reliability, QAMS can be applied for quality evaluation of Huangzhi Neiyi Capsules.

Study on Quality Standard of Yiwei Xiaoyu Granules / 中国中医药信息杂志

Objective To establish the quality standard for Yiwei Xiaoyu Granules.Methods TLC was used for the qualitative analyses of Atractylodis Macrocephalae Rhizoma, Angelicae Sinensis Radix and Dendrobii Caulis; HPLC was applied to determine the contents of notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1and ginsenoside Re. Results The TLC spots were clear and free from negative interference. Notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1and ginsenoside Re showed good linear relationships within the ranges of 0.067 2–0.672 μg (r=0.999 6), 0.501 2–5.012 μg (r=0.999 6), 0.326 5–3.265 μg (r=0.999 6), 0.098 3–0.983 μg (r=0.999 7), whose average recoveries were 96.32% (RSD=1.3%), 96.45% (RSD=1.5%), 101.23% (RSD=1.7%), and 97.89% (RSD=1.7%), respectively. Conclusion This method is accurate, reliable, and reproducible, which can be used for the quality control of Yiwei Xiaoyu Granules.

Study on Optimization of the Extraction Technology of Bushen Yangxue Granules / 中国药房

OBJECTIVE: To optimize the extraction technology of Bushen yangxue granules, and to provide reference for developing hospital preparations. METHODS: The orthogonal test was designed to optimize the extraction technology of Bushen yangxue granules using the comprehensive score including the content of hyperoside and total flavonoids, extract yield as evaluation indexes, water amount, extraction time and extraction times as factors. Verification test was also conducted. RESULTS: The extraction technology of Bushen yangxue granules was 10-fold water, extracting for 2 times and 1 hour each time. The results of verification test indicated that the contents of hyperoside and total flavonoids were 0. 154 mg/g (RSD=0. 010％, n=3) and 11. 094 mg/g (RSD=0. 008％, n=3). The average extract yield was 37. 76％ (RSD=0. 008％, n=3). CONCLUSIONS: The optimized extraction technology is stable and feasible, which can provide the basis for developing hospital preparations of Bushen yangxue granules.

Application of ETDRS chart and mfERG in assessing pathologic myopia combined with macular CNV after treatment / 国际眼科杂志(Guoji Yanke Zazhi)

·AIM: To evaluate the efficacy of anti - vascular endothelial growth factor ( VEGF ) and photodynamic therapy ( PDT) on pathological myopia ( PM) combined with choroidal neovascularization ( CNV ) using ETDRS chart and multifocal electroretinogram (mfERG). ·METHODS: Forty-three patients ( 45 eyes) diagnosed by fundus fluorescein angiography ( FFA ), indocyanine green angiograph ( ICGA ) and optical coherence tomography (OCT) with PM combined with macular CNV were recruited in this study. The patients were randomly divided into two groups for different treatments, intravitreal injection with Ranibizumab ( 20 patients, 22 eyes) and PDT (23 patients, 23 eyes). After treatment, all patients had been followed up monthly for 12mo. The further treatments were operated according to referral situations. The best corrected visual acuity ( BCVA) was recorded with the ETDRS chart and mfERG. At the last follow-up, the therapy efficacy was determined by ETDRS numbers and mfERG and analyzed. ·RESULTS: Before treatment, there was no significant difference on the baseline in ETDRS and mfERG latency of N1 wave, latency and the density values of P1 wave between ranibizumab group and PDT group. After 12mo treatment, the ETDRS number in ranibizumab group (39. 23± 20. 06) significantly increased to the baseline by 5. 88±9. 03(P<0. 05), and in PDT group (37. 38±16. 95) was not significantly improved by 0. 33±6. 94(P>0. 05). There was no significant difference in latency of N1 wave, latency and the density values of P1 wave from treatment response of mfERG in the two groups (P>0. 05). · CONCLUSION: In the treatment of macular CNV complicated by the PM, ranibizumab injection can improve visual function better than PDT, while similar on macular.

Evaluation of Ranibizumab and PDT for patients with pathologic myopia and macular CNV / 国际眼科杂志(Guoji Yanke Zazhi)

·AIM: To evaluate the efficacy of anti - vascular endothelial growth factor (VEGF) and photodynamic therapy (PDT) on pathological myopia (PM) combined with choroidal neovascularization (CNV). ·METHODS: Forty-three patients (45 eyes) diagnosed by fundus fluorescein angiography (FFA), indocyanine green angiography ( ICGA ) and optical coherence tomography (OCT) with PM combined with macular CNV were recruited in this study. The patients were randomly divided into two groups for different treatments, intravitreal injection with Ranibizumab (20 patients, 22 eyes) and PDT(23 patients,23 eyes). After treatment,all patients had been followed up monthly for 12mo. The further treatments were operated according to referral situations. The best corrected visual acuity (BCVA) was recorded with the ETDRS chart and the mean defect(MD) of the center 10° visual field was measured. At the last follow-up,the therapy efficacy was determined by ETDRS numbers and MD and analyzed. ·RESULTS: Before treatment, there was no significant difference on the baseline in ETDRS and MD between ranibizumab group and PDT group (P>0.05). After 12mo treatment, the ETDRS number in ranibizumab group (39.23±20.06) significantly increased (by 5.88 ± 9.03, P<0.05), but the PDT group (37. 38 ± 16. 95) was not significantly improved(by 0.33±6.94,P>0.05). The MD in ranibizumab group decreased significantly (P<0.05), and no significant change was found in PDT group(P>0.05). · CONCLUSION: In the treatment of macular CNV complicated by the PM, ranibizumab injection can improve visual function better than PDT.

Periostin promotes migration and invasion of renal cell carcinoma through the integrin/focal adhesion kinase/c-Jun N-terminal kinase pathway.

Periostin (POSTN) is an extracellular matrix protein which is overexpressed in a variety of cancers and has been related to tumorigenesis of renal cell carcinoma. However, the involvement of POSTN in renal cell carcinoma migration, invasion, and their underlying mechanisms has not been established. In this study, renal cell carcinoma cell lines stably overexpressing POSTN were established using a lentiviral vector, and the effects of POSTN on renal cell carcinoma cell migration and invasion were investigated. POSTN overexpression increased the migration and invasion capabilities of renal cell carcinoma cell lines as well as activity of matrix metalloproteinase-2 and matrix metalloproteinase-9. Integrin αvß3 and αvß5 antibodies inhibited POSTN overexpression or recombinant POSTN-induced focal adhesion kinase activation, cell migration, and invasion. Furthermore, lentivirus-mediated focal adhesion kinase knockdown and c-Jun N-terminal kinase inhibitor reduced POSTN-enhanced phosphorylation of c-Jun N-terminal kinase, matrix metalloproteinase-9 and matrix metalloproteinase-2 expressions, cell migration, and invasion. Our research thus indicates that POSTN promotes renal cell carcinoma cell migration and invasion through interaction with integrins αvß3 and αvß5 and subsequent activation of the focal adhesion kinase/c-Jun N-terminal kinase pathway. These results suggest that POSTN plays a critical role in renal cell carcinoma metastasis and may represent a potential target for novel therapeutic approaches against renal cell carcinoma.

Inwardly rectifying K+ channels are major contributors to flow-induced vasodilatation in resistance arteries.

KEY POINTS: Endothelial inwardly rectifying K+ (Kir2.1) channels regulate flow-induced vasodilatation via nitric oxide (NO) in mouse mesenteric resistance arteries. Deficiency of Kir2.1 channels results in elevated blood pressure and increased vascular resistance. Flow-induced vasodilatation in human resistance arteries is also regulated by inwardly rectifying K+ channels. This study presents the first direct evidence that Kir channels play a critical role in physiological endothelial responses to flow. ABSTRACT: Inwardly rectifying K+ (Kir) channels are known to be sensitive to flow, but their role in flow-induced endothelial responses is not known. The goal of this study is to establish the role of Kir channels in flow-induced vasodilatation and to provide first insights into the mechanisms responsible for Kir signalling in this process. First, we establish that primary endothelial cells isolated from murine mesenteric arteries express functional Kir2.1 channels sensitive to shear stress. Then, using the Kir2.1+/- heterozygous mouse model, we establish that downregulation of Kir2.1 results in significant decrease in shear-activated Kir currents and inhibition of endothelium-dependent flow-induced vasodilatation (FIV) assayed in pressurized mesenteric arteries pre-constricted with endothelin-1. Deficiency in Kir2.1 also results in the loss of flow-induced phosphorylation of eNOS and Akt, as well as inhibition of NO generation. All the effects are fully rescued by endothelial cell (EC)-specific overexpression of Kir2.1. A component of FIV that is Kir independent is abrogated by blocking Ca2+ -sensitive K+ channels. Kir2.1 has no effect on endothelium-independent and K+ -induced vasodilatation in denuded arteries. Kir2.1+/- mice also show increased mean blood pressure measured by carotid artery cannulation and increased microvascular resistance measured using a tail-cuff. Importantly, blocking Kir channels also inhibits flow-induced vasodilatation in human subcutaneous adipose microvessels. Endothelial Kir channels contribute to FIV of mouse mesenteric arteries via an NO-dependent mechanism, whereas Ca2+ -sensitive K+ channels mediate FIV via an NO-independent pathway. Kir2 channels also regulate vascular resistance and blood pressure. Finally, Kir channels also contribute to FIV in human subcutaneous microvessels.

Changes of choroidal thickness after ranibizumab treatment for non-proliferative diabetic retinopathy with macular edema / 眼科新进展

Objective To observe the changes of choroidal thickness after ranibizumab treatment for non-proliferative diabetic retinopathy (NPDR) with macular edema,and determine the association between choroidal thickness and visual acuity.Methods Twenty-three eyes from 23 patients diagnosed with NPDR and diabetic macuiar edema were treated with 3 monthly intravitreal injections of ranibizumab.The subfoveal choroidal thickness and central macular thickness were measured,and the best corrected visual acuity was recorded.Changes of subfoveal choroidal thickness,correlation between subfoveal choroidal thickness and best corrected visual acuity were assessed at 3 months follow-up.Results After 3 monthly anti-VEGF treatments,subfoveal choroidal thickness and central macular thickness decreased significantly,there was no statistical difference at 1 month and 2 months compared with pre-treatment (all P ＞ 0.05),but there was statistical differences at 3 months compared with pre-treatment (P =0.04,0.01).In the treatment,the best corrected visual acuity increased gradually,there was no statistical difference at 1 month and 2 months compared with pre-treatment (all P ＞ 0.05),but there was statistical differences at 3 months compared with pre-treatment (P =0.04).Before the treatment,the subfoveal choroidal thickness was positive correlated with macuiar retinal thickness (R2 =0.94,P =0.00);And the best corrected visual acuity before treatment and 3 months after treatment had a positive correlation (R2 =0.93,P =0.00;R2 =0.82,P =0.00).There was a positive correlation between central macular thickness and best corrected visual acuity at 3 months after treatment (R2 =0.83,P =0.00).There was a positive correlation in the best corrected visual acuity between before treatment and 3 months after treatment (R2 =0.84,P =0.00).Conclusion The subfoveal choroidal thickness is a clinical index to evaluate the efficacy of anti-VEGF in the treatment of diabetic macuiar edema.Baseline subfoveal choroidal thickness can be considered as one of the indicators of clinical efficacy in the short term.

High-throughput identification of novel conotoxins from the Chinese tubular cone snail (Conus betulinus) by multi-transcriptome sequencing.

BACKGROUND: The venom of predatory marine cone snails mainly contains a diverse array of unique bioactive peptides commonly referred to as conopeptides or conotoxins. These peptides have proven to be valuable pharmacological probes and potential drugs because of their high specificity and affinity to important ion channels, receptors and transporters of the nervous system. Most previous studies have focused specifically on the conopeptides from piscivorous and molluscivorous cone snails, but little attention has been devoted to the dominant vermivorous species. RESULTS: The vermivorous Chinese tubular cone snail, Conus betulinus, is the dominant Conus species inhabiting the South China Sea. The transcriptomes of venom ducts and venom bulbs from a variety of specimens of this species were sequenced using both next-generation sequencing and traditional Sanger sequencing technologies, resulting in the identification of a total of 215 distinct conopeptides. Among these, 183 were novel conopeptides, including nine new superfamilies. It appeared that most of the identified conopeptides were synthesized in the venom duct, while a handful of conopeptides were identified only in the venom bulb and at very low levels. CONCLUSIONS: We identified 215 unique putative conopeptide transcripts from the combination of five transcriptomes and one EST sequencing dataset. Variation in conopeptides from different specimens of C. betulinus was observed, which suggested the presence of intraspecific variability in toxin production at the genetic level. These novel conopeptides provide a potentially fertile resource for the development of new pharmaceuticals, and a pathway for the discovery of new conotoxins.

Relationship of long noncoding RNA and viruses.

Long noncoding (lnc)RNAs comprise a diverse group of transcripts including large intervening noncoding (linc)RNAs, natural antisense transcripts (NATs) and intronic lncRNAs. The functions and mechanisms of more than 200 lncRNAs have been studied in vitro and the results suggest that lncRNAs may be molecular markers of prognosis in cancer patients. Some lncRNAs can promote virus replication and allow escape from cytosolic surveillance to suppress antiviral immunity. For example, lncRNA can cause persistent infection by Theiler's virus, and microRNA (miR)-27a/b is important for efficient murine cytomegalovirus (MCMV) replication. The available evidence suggests that lncRNAs may be potential targets of novel antiviral drugs.

Suppression of wingless-type MMTV integration site family, member 1 expression by small interfering RNA inhibits U251 glioma cell growth in vitro.

A Wingless-type MMTV integration site family, member 1 (Wnt-1) RNA interference expression vector was constructed during the present study, which was used to transfect the glioma U251 cell line and investigate its effect on glioma. Two 21-base oligonucleotides complementary to the coding sequence that was flanking the loop sequence were designed to form a DNA hairpin template for the target small interfering RNA (siRNA). The siRNA templates were cloned into the siRNA expression vector, pGPU6/green fluorescent protein (GFP)/Neo and the sequence was confirmed by DNA sequencing. The pGPU6/GFP/Neo-short hairpin RNA (shRNA)-Wnt-1 vector was subsequently transfected into U251 cells, and reverse transcription polymerase chain reaction and western blot analysis were used to evaluate the Wnt-1 gene silencing effect on U251 cell growth by MTT assay and flow cytometry. The Wnt-1 protein expression was significantly reduced following transfection with the recombinant plasmid, as determined by western blot analysis of the transfected U251 cells. This transfection exhibited a significantly higher death rate, as shown by MTT. Thus, the present study demonstrated that the pGPU6/GFP/Neo-shRNA-Wnt-1 vector inhibited Wnt-1 protein expression. However, further investigations regarding the Wnt signaling pathway in glioma pathogenesis are required.