Objective To explore the role of long noncoding RNA ( lncRNA) CTD-2012K14. 6 in the development of gestational diabetes mellitus (GDM) related macrosomia. Methods The quantitative real-time PCR ( qRT-PCR) was performed to measure the expression of CTD-2012K14.6 in placentas of women with or without GDM, and the quantity of CTD-2012K14. 6 expression and its association with fetal weights were analyzed; Bioinformatic analysis was performed to predict the downstream molecules. CTD-2012K14. 6 over-expressing lentiviral and siRNA was constructed in human trophoblastic cell line HTR-8/SVneo cells, qRT-PCR and Western blot (WB) were used to invest its effect in modulating the expression of downstream molecules. Results The expression of CTD-2012K14.6 in GDM placentas was significantly higher than that in normal controls (1.70 ± 0.63 vs 1.00 ± 0.56,t=3.68,P＜0.01), and positively correlated with fetal weight (r=0.8501, P＜0.01); on-line analysis showed that CTD-2012K14.6 was located at chr16:67,549,214-67,563,958, which was located in the intron of CCCTC-binding factor( CTCF); Up-regulating CTD-2012K14.6 could significantly reduce the expression of CTCF mRNA and protein, and increase the expression of insulin-like growth factor-Ⅱ( IGF-Ⅱ) mRNA and protein, while down-regulating CTD-2012K14.6 could significantly increase the expression of CTCF mRNA and protein, and reduce the expression of IGF-ⅡmRNA and protein. Conclusion The CTD-2012K14. 6 may play an important role in the pathogenesis of GDM related macrosomia by upregulating the expression of CTCF and IGF-Ⅱ.

To establish a method for determination contents of schizandrin, tanshinone I, cryptotanshinone, tanshinone II (A) and schizandrin B in rongxin pills. The HPLC method was performed on an Agilent C18. The mobile phase was composed of methnol and water wish gradient elution. The flow rate was 1.0 mL x min(-1). The column temperature was 30 degrees C and the detection wavelength wash 240 nm. The linear of schizandrin, tanshinone I, cryptotanshinone, Tanshinone II (A) and schizandrin B were 3.000-48.00 (r = 1.000), 3.985-63.76 (r = 0.999 9), 6.370-101.9 (r = 1.000), 8.690-139.0 (r = 0.999 9), 1.700-27.20 mg x L(-1) (r = 0.999 9), respectively. The average recoveries were 98.44%, 100.3%, 99.29%, 99.07%, 98.42%, and RSDs were 0.61%, 1.1%, 0.52%, 0.72%, 0.97%. The method is convenient, accurate and has good precision. It can be used for determination of the preparation.
Chromatography, High Pressure Liquid ; Cyclooctanes ; analysis ; Diterpenes, Abietane ; analysis ; Drugs, Chinese Herbal ; chemistry ; Lignans ; analysis ; Linear Models ; Organic Chemicals ; analysis ; Phenanthrenes ; analysis ; Polycyclic Compounds ; analysis ; Quality Control ; Reproducibility of Results

Acute Disease ; Administration, Inhalation ; Animals ; Disease Models, Animal ; Hypertension, Pulmonary ; drug therapy ; Male ; Nitric Oxide ; administration & dosage ; therapeutic use ; Nitrites ; administration & dosage ; therapeutic use ; Rats ; Rats, Sprague-Dawley

Objective To explore the value of transrectal ultrasound (TRS) in diagnosing middle ureteral calculi.Methods Patients who were suspected as middle urinary calculi by clinicians underwent transabdominal ultrasound (TAS) and transrectal ultrasound in the First Affiliated Hospital of Guangzhou Medical University and Guangzhou Hospital of Integrated Traditional and West Medicine from 2011 to 2012. The diagnosis were finally confirmed by a variety of methods .Results Forty patients were eventually diagnosed as middle ureteral calculi by X-ray,computer tomography (CT),surgery,intravenous pyelography (IVP)imaging and magnetic resonance imaging (MRI).The other 5 cases were diagnosed as ureteral tumor , ureteral cystis(confirmed by MRI)and oppressed ureteral by gynecology cystis (confirmed by surgery).TRS detected 37 of 40 middle ureteral calculi(92.5%).TAS detected 8 of 40 middle ureteral calculi(20.0%). The detection ratio of TRS was higher than that of TAS and the difference was statistically significant (P=0.00).TAS could display ureter across iliac blood vessels expansion in all 40 patients.Conclusions Using TRS in diagnosing middle ureteric calculi is quick , convenient with high sensitivity and worthy of wide application.TAS can display ureter across iliac blood vessels expansion and can help to diagnosis of middle ureteral calculi.

Objective To investigate the difference of expression profiling of plasma miRNAs (microRNA) between the patients with small artery occlusive stroke (SAO) and the healthy subjects.Methods Eight patients with SAO classified by TOAST were selected and 8 healthy subjects were used as a control group.High-throughput sequencing technology was used to detect the expression profiling of plasma miRNAs.The differentially expressed miRNAs were screened.Real-time fluorescent quantitative polymerase chain reaction was used to validate the results,and the target gene prediction and bioinformatics analysis were performed.Results The miRNA difference analysis showed that the expression profilings of miRNA-127,miRNA-99b-5p,miRNA-320,and other 19 miRNAs in the SAO group were significantly upregulated compared with those in the control group (all P＜0.01),while miRNA-451a and other 5 miRNAs in the SAO group were significantly downregulated compared with those in the control group (all P ＜0.01).The validated results of miRNA-127,miRNA-99b-5p,miRNA-320,and miRNA-451a with real-time fluorescent quantitative polymerase chain reaction were consistent with those of the high-throughput sequencing.Bioinformatics analysis showed that the miRNA-regulated target genes expressed differentially were mainly correlated with cell proliferation,adhesion,phylogenetic development,macromolecule metabolism,and other biological functions.Conclusions There are significant differences in the expression profiling of plasma miRNAs between the patients with SAO and the healthy subjects,suggesting that miRNAs may play a regulatory role via target genes in pathogenesis of SAO.

Objective To investigate the relationship between bed rest time and development of prethrombotic state in the elderly patients with hip fracture.Methods One hundred and sixty-six patients who stayed in bed after hip fracture,aged ≥65 yr,were divided into 5 groups according to the bed rest time on admission to hospital:bed rest time ＜ 24 h (Ⅰ group,n =61),bed rest time 24-48 h (Ⅱ group,n =29),bed rest time 3-6days (Ⅲ group,n =29),bed rest time 7-14 days (Ⅳ group,n =34),and bed rest time ＞ 14 days (Ⅴ group,n =13).Venous blood samples were collected to measure the platelet count,coagulation function,and concentrations of plasma D-Dimer and serum α-granule membrane protein 140 (GMP-140).The development of abnormality in each index was recorded.The development of deep vein thrombosis in both lower extremities was diagnosed using color Doppler ultrasound in D-Dimer-positive patients.Results Compared with group Ⅰ,the abnormal rate of fibrinogen (Fib) and D-Dimer and serum GMP-140 concentrations were significantly increased in Ⅱ and Ⅲ groups,the abnormal rate of platelet count,Fib and D-Dimer and serum GMP-140 concentrations were increased in lⅣ group,and the abnormal rate of platelet count was increased,and no significant change was found in the serum GMP-140 concentrations and abnormal rate of Fib and D-Dimer in Ⅴ group.Compared with group Ⅱ,the serum GMP-140 concentrations were significantly increased in Ⅲ and Ⅳ groups,the abnormal rate of Fib and D-Dimer was increased in Ⅳ group,and no significant change was found in the abnormal rate of Fib and DDimer in Ⅲ group.The abnormal rate of platelet count was significantly lower in Ⅳ group than in Ⅲ group.Color Doppler Ultrasonography showed no sign of deep vein thrombosis.Conclusion For the elderly patients with hip fracture,the possibility of pre-thrombotic state developed is increased when the bed rest time is more than 24 h,and the patients were classified as high-risk patients when the bed rest time is more than 3 days.

BACKGROUND: Few studies have reported the effect of aldosterone receptor antagonist (ARA) on myocardial remodeling after acute myocardial infarction (AMI). This study was undertaken to investigate the preventive effect of ARA on myocardial remodeling after AMI.METHODS: A total of 616 patients who had been admitted into the CCU of the First Affiliated Hospital of Harbin Medical University from January 2008 to January 2010 were studied prospectively. Only 528 patients were observed completely, including 266 of the control group and 262 of the treatment group. There was no statistical difference in age, gender, medical history, admission situation, and treatment between the two groups (P>0.05). The preventive effects of spironolactone on cardiac remodeling, left ventricular function, renal function and blood levels of potassium were evaluated by echocardiography, serum potassium and serum creatinine at one-month and one-year follow-up.RESULTS: The echocardiography indicators such as LVESD, LVEDD, LVEF, LAD-ML and LAD-SI were signifi cantly improved in the treatment group compared with the control group at one year (P<0.05). In the treatment group, LVESD, LVEDD, LVPWT, LVEF, LAD-ML and LAD-SI were more significantly improved at one year than one month (P<0.05,P=0.007 to LVEF), and in the control group LVEF was more signifi cantly improved at one year than one month (P=0.0277). There were no signifi cant differences in serum potassium and serum creatinine levels between the two groups.CONCLUSION: On the basis of conventional treatment, the early combination of low-dose spironolactone (20 mg/d) could inhibit cardiac remodeling at late stage and prevent heart failure.

OBJECTIVETo compare the difference in clinical efficacy on polycystic ovary syndrome (PCOS) between electroacupuncture (EA) and dyne-35 and to explore the effect mechanism.

METHODSSixty-five patients were randomized into an EA group (33 cases) and a western medication group (32 cases). In the EA group, the selected acupoints were Danzhong (CV 17), Qimen (LR 14), Zhongwan (CV 12), Tianshu (ST 25), Guanyuan (CV 4), Zigong (EX-CA 1), Sanyinjiao (SP 6), Zusanli (ST 36) and Taichong (LR 3), etc. After the arrival of qi, electric stimulation was attached to the acupoints for 30 min. The treatment was given 3 times a week. In the western medication group, dyne-35 was prescribed on the 5th day of natural menstruation or withdrawal bleeding, one tablet a day, continuously for 21 days. The treatment cycle was 3 months in the two groups. The menstrual condition, body mass, body mass index (BMI), serum testosterone (T), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and LH/FSH were compared before and after treatment in the two groups. The clinical efficacy was assessed in the two groups.

RESULTSThe total effective rate was 90.6% (29/32) in the EA group and was 93.3% (28/30) in the western medication group. The efficacy was similar in the two groups (P > 0.05). After treatment, the levels of LH and LH/FSH were all reduced significantly in the two groups (all P < 0.01). After treatment, T level in serum was reduced apparently in the western medication group (P < 0.05). Before and after treatment, the differences in body mass and BMI in the EA group were more significant than those in the western medication group (P < 0.01, P < 0.05).

CONCLUSIONEA is the effective method for PCOS, similar to that of dyne-35. The effect of it for weight loss is superior to dyne-35 and no apparent adverse reactions happen. The effect mechanism of EA is related to the regulation of serum sexual hormone levels and their ratio, as well as to the regulation of body lipid metabolism.

Acupuncture Points ; Adolescent ; Adult ; Cyproterone Acetate ; administration & dosage ; Drug Combinations ; Electroacupuncture ; Ethinyl Estradiol ; administration & dosage ; Female ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Polycystic Ovary Syndrome ; blood ; drug therapy ; therapy ; Young Adult

Objective To compare the relative efficacy and quality of extraction of human fecal DNA using four methods.Methods Real-time PCR were utilized for analysis both quantification and quality of the fecal targeted bacteria(including gut all eubaeterium,Bacteriodes-PrevoteUa group,Bifidobacterium spp Enterobacteriaceae and Enterococcus spp)by using 16s rRNA gene-targeted genus or group-specific primer sets.Results The negative rat of PCR product from method 3(phenol-chloroform plus bead-beating) was about 40%(4/10)by using universal primers,the PCR inhibition disappeared after fecal DNA purified with column.The total fecal 16s rRNA gene copy numbers(per gram of wet weight of feces)as well as the numbers of Bacteriodes-Prevotella group from method 1(QIAamp~DNA stool mini kit)and 4(QIAamp~ DNA stool mini kit combined with bead-beating)was higher significantly than that from method 2(FastDNA ~Kit,Biol01)and 3(P

OBJECTIVETo investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD).

METHODSThe levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aβ25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again.

RESULTSCompared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aβ25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aβ25-35. Compared to controls, the Aβ25-35-treated SH-SY5Y cells and the Aβ25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former.

CONCLUSIONThe siRNA silencing of α3 nAChR mRNA may enhance the effect of Aβ25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.

Alzheimer Disease ; etiology ; Amyloid beta-Peptides ; metabolism ; Apoptosis ; Cell Line, Tumor ; Gene Silencing ; Humans ; Neuroblastoma ; metabolism ; pathology ; Peptide Fragments ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Nicotinic ; genetics ; metabolism ; Signal Transduction ; Transfection ; bcl-2-Associated X Protein ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism