Objective To investigate the impact of propofol(Pro)on high glucose(HG)-induced myocardial cell injury through autophagy mediated by forkhead box O1(Fox O1)/thioredoxininteracting protein(TXNIP)signaling pathway.Methods H9c2 cells were divided into blank group(5.5 mmol·L-1 glucose),high glucose(HG)group(30 mmol·L-1 glucose),HG+Pro group(30 mmol·L-1 glucose+25 μmol·L-1 Pro),HG+Pro+negative control(OE NC)group(first transfected with OE NC,then treated with 30 mmol·L-1 glucose+25 μmol·L-1 Pro),HG+Pro+Fox O1 overexpression plasmid(Fox O1-OE)group(first transfected with Fox O1-OE,then treated with 30 mmol·L-1 glucose+25 μmol·L-1 Pro).Cell counting kit-8(CCK-8)method,TdT-mediated dUTP nick end labeling(TUNEL)method,enzyme-linked immunosorbent assay(ELISA),transmission electron microscope and Western blot were applied to detect the cell survival rate,apoptosis rate,superoxide dismutase(SOD)and malondialdehyde(MDA)levels in the supernatant,and the changes in Autophagosome,Fox O1/TXNIP and autophagy protein expression levels.Results The cell viabilities of blank group,HG group,HG+Pro group,HG+Pro+OE NC group and HG+Pro+Fox O1-OE group were(100.00±0.00)％,(48.15±4.82)％,(79.66±7.98)％,(78.89±7.91)％and(49.22±4.93)％,respectively;the apoptosis rates were(12.04±1.21)％,(42.34±4.25)％,(26.22±2.63)％,(27.02±2.71)％,(38.29±3.86)％,respectively;SOD levels were(62.24±6.25),(28.21±2.85),(55.37±5.58),(55.09±5.53),(30.66±3.08)U·mg-1 pro,respectively;MDA levels were(0.38±0.04),(1.43±0.15),(0.67±0.07),(0.72±0.08)and(1.34±0.14)U·mg-1 pro,respectively;the number of autophagosomes was 6.24±0.63,13.05±1.32,8.31±0.84,8.55±0.86 and 12.22±1.23,respectively;phosphorylated Fox O1(p-Fox O1)/Fox O1 ratios were expressed as 0.34±0.04,0.86±0.09,0.48±0.05,0.43±0.05 and 0.74±0.08;TXNIP were expressed as 0.24±0.03,0.62±0.08,0.38±0.04,0.36±0.04 and 0.58±0.06;Bcelin-1 were expressed as 1.12±0.12,0.53±0.06,1.02±0.11,1.05±0.11 and 0.62±0.07;p62 were expressed as 0.56±0.06,1.56±0.16,0.82±0.09,0.86±0.09 and 1.44±0.15;there were statistical differences in the above indexes between HG group and blank group,HG+Pro group and HG group,HG+Pro+Fox O1-OE group and HG+Pro+OE NC group(all P＜0.05).Conclusion Pro inhibits autophagy by inhibiting Fox O1/TXNIP signaling pathway,and improves HG-induced myocardial cell injury.

Objective: To construct the mmu-miR-155 eukaryotic overexpression vector pmR-155 and to investigate its effect on HBV replication and expression of PTEN in vivo. Methods: The mmu-mir-146a precursor gene fragment pre-mmu-mir-146a was amplified by PCR, then connected to the pmR-mCherry plasmid vector after double enzyme digestion, the accuracy of recombinant vector was verified by colony PCR、double enzyme digestion and sequencing; then the recombinant vector was transfected HBV transgene mice(Experimental Group)with hydrodynamics-based injection via vena caudalis, and pmR-mCherry plasmid、PBS were respectively transfected into the mice as Empty plasmid Group、Blank Group. The concentration of IFN-γ in the serum was detected by ELISA. The expression of SOCS1、PTEN mRNA in the liver was detected by qPCR at 30d post-transfectioned. The Western blot was performed to detect the changes in SOCS1、PTEN、HBX in the liver tissue at 30 d post-transfectioned. The results were analyzed with Student’s t-test, or one-way analysis of variance and the least significant difference test. Results: the colony PCR、double enzyme digestion and sequencing verified that the gene was inserted into the pmR-mCherry vector. Compared with Blank Group, the expression of miR-155 in the Experimental Group was significantly increased(t = 8.90, P < 0.01); the concentration of IFN-γ in the Experimental Group was significantly increased(F = 26.58, P < 0.01); the mRNA(F_{SOCS1 mRNA} = 19.72, P < 0.01; F_{PTEN mRNA} = 7.38, P < 0.05) and protein(F_SOCS1 = 50.30, P < 0.01; F_PTEN = 129.00, P < 0.01) expression of COCS1、PTEN was significantly decreased in the Experimental group and the protein of HBX was also significantly(F_HBX = 77.97, P < 0.01). Conclusion The pmR-155 eukaryotic overexpression vector is successfully constructed, this recombinant vector can express miR-155 stably; miR-155 can down-regulate cocs1、PTEN gene expression and up-regulate the expression of IFN-γ, it can inhibit the replication of HBV and a potential targets to treating hepatocellular carcinoma.

Objective To construct the has-miR 146a eukaryotic overexpression vector pmR 146a and to explore its effect on the expression of c-Myc gene in HepG2.2.15 cells.Methods The has-miR-146a precursor gene fragment pre-has-miR-146a was amplified by PCR,then connected to the pmR-mCherry plasmid vector after double enzyme digestion,the accuracy of recombinant vector was verified by colony PCR,double enzyme digestion and sequencing;then the recombinant vector was transfected into HepG2.2.15 cells as the experimental group,meanwhile the empty vector group (transfecting pmR-mCherry empty plasmid group) and blank group(transfecting reagent lip2000+PBS),then the fluorescent protein expression amount was observed under the fluorescence microscopy at 24,48 h;the expression of has miR-146a was evaluated by qPCR;at 24,48 h after transfection,the expression levels of c-Myc gene mRNA were detected by qPCR,and the c-Myc protein expression level after 48 h was detected by Western blot.Results The colony PCR,double enzyme digestion and sequencing verified that the pre-has-miR-146a gene fragment was inserted into the pmR-mCherry vector;at 24,48 h after transfection in the experimental group and empty vector group,intracellular strong fluorescence was seen by fluorescent microscope,the transfection efficiency was at 50％-60％ contrasting without fluorescence;the has-miR-146a expression level in the experimental group was significantly higher than that in the empty vector group and blank group (P＜0.01);the c-Myc mRNA expression at 24,48 h after tranfection was significantly lower than that in the empty vector group and blank group (P＜0.05);the protein expression amount at 48 h after transfection was lower than that in the empty vector group and blank group (P＜0.01).Conclusion The pmR-146a eukaryotic overexpression vector is successfully constructed,this recombinant vector can express miR-146a stably;miR-146a can down-regulate c-Myc cancer gene expression,which can serve as one of potential targets for treating hepatocellular carcinoma.

Abelmoschus manihot (L.) Medic., a folk herbal medicine in China, is a flowering plant belonging to Abelmoschus L. genus and Malvaceae family, which has been reported with an antidepressant activity. The study was designed to isolate flavonoids from Abelmoschus manihot corolla and explore the action mechanism of antidepressant activities. The flavonoids were isolated and purified by D101 macroporous resin column, polyamide column and Sephadex LH-20 sequentially and identified as myricetin-3-O-β-D-glucoside (1), gossypetin-8-O-β-D-glucuronide (2, G-8-G), gossypetin-3'-O-β-D-glucoside (3), quercetin-3'-glucoside (4, Q-3-G), isoquercitrin (5, IQT), hyperoside (6, HY), myricetin (7), quercetin (8, QT). Compounds 2, 4, 5, 6 and 8 (15, 30 and 60 mg·kg^-1) were orally administered to mice and the reaction was observed in tail suspension test (TST) and forced swimming test (FST). Western blot analysis was used in determination of the protein expressions of brain-derived neurotrophic factor (BDNF), tyrosine receptor kinase B (TrkB) and phosphorylation eukaryotic elongation factor 2 (p-eEF2). The results revealed that only Q-3-G and G-8-G (15, 30, 60 mg·kg^-1) significantly reduced the immobility time in FST and TST. Furthermore, Q-3-G and G-8-G remarkably increased the expression of BDNF and TrkB, and decreased the expression of p-eEF2. These results suggest that Q-3-G and G-8-G had an obvious antidepressant activity via up-regulation of BDNF expression. The new observation will provide a new direction in the development of antidepressant in the treatment of major depressive disorder (MDD).

Objective To construct the miRNA‐21 eukaryotic overexpression vector pmR‐21 and to explore its regulation effect on the expression of c‐myc gene in HepG2 .2 .15 cells .Methods The miRNA‐21 precursor gene fragment pre‐miRNA‐21 was amplified by PCR ,then connected to the pmR‐mCherry plasmid vector after double enzyme digestion ,the accuracy of the recombi‐nant vector was verified by double enzyme digestion and sequencing ;then the recombinant vector was transfected into HepG2 .2 .15 cells ,the fluorescent protein expression was observed under the fluorescence microscopy at 24 h and the transfection efficiency was detected by flow cytometry ;the expression of miRNA‐21 was evaluated by real‐time quantitative PCR;at 72 h after transfection ,the expression levels of c‐myc gene were detected by RT‐PCR and Western blot ;CCK‐8 was used to detect the cell proliferation in each group .Results The double enzyme digestion and Western blot verified that the target gene fragment was inserted into the pmR‐mCherry vector;at 24 h after transfection ,intracellular strong fluorescence was seen ,the transfection efficiency was higher than 50% ;miRNA‐21 expression level of the pmR‐21 recombinant vector group was significantly increased;c‐myc gene expression was increased in the pmR‐21 recombinant vector group at 72 h after transfection ,the cell proliferation in the pmR‐21 recombinant group was faster than that in the control group(P<0 .05) .Conclusion The pmR‐21 eukaryotic overexpression vector is successfully con‐structed ,this recombinant vector can express miRNA‐21 stably ;miRNA‐21 can up‐regulate c‐myc gene expression ,c‐myc gene is one of miR‐21′s targets for playing a cancer‐promoting action .

Objective To study the effect of early mechanical ventilation in treatment of patients with severe acute pancreatitis(SAP).Methods Fifty-two patients with SAP admitted in the First People's Hospital of Zhaoqing from January 2010 to January 2015 were randomly allocated into two groups (n =26),early mechanical ventilation group(observation group) and conventional mechanical ventilation group(control group).Patients in the observation group treated with early lung protective ventilation when PaO2 ＜ 13.3kPa.Patients in the control group treated without mechanical ventilation untill PaO2 ＜ 8kPa.The symptoms,the extent of inflammatory reaction,the severity of lung lesions and the mortality of two groups were compared through monitoring vital signs,abdominal circumference,APACHE Ⅱ score,bladder pressure,oxygenation index (PaO2/FiO2),C reactive protein (CRP),procalcitonin (PCT),hospital stay and mortality.Results No statistically significant differences in the APACHE Ⅱ score,bladder pressure,oxygenation index,CRP and PCT in two groups before treatment were observed(P ＞ 0.05).The APACHE Ⅱ score (12.8 ± 7.6) points,bladder pressure (14.9± 7.9) cmH2O,CRP (48.8 ± 30.1) rmg/L,PCT (1.25 ± 0.55) μg/L,mortality (3.84％) of the observation group after treatment were significantly lower than those of the control group (t =2.057,2.091,3.252,2.697,x2 =4.305,all P ＜ 0.05),while the oxygenation index in the observation group [(300.0 ± 34.9) mmHg] was significantly higher than that in the control group [(278.1 ± 32.8) mmHg],the difference of the two groups was statistically significant (t =3.322,P ＜ 0.05).Conclusion Early lung protective ventilation is safe and effective for treatment of the patients with SAP.

OBJECTIVETo investigate the correlation between signal transducer and activator of transcription 3 (STAT3) expression and pituitary adenoma subtypes.

METHODThe STAT3 expression profiles in different pituitary adenomas from 74 patients were determined using quantificational real-time polymerase chain reaction,Western blot,and immunohistochemistry.

RESULTSExpression of STAT3 was observed in all pituitary adenoma subtypes. The STAT3 expression level was highest in growth hormone adenoma when compared with other tumors including prolactin,follicle-stimulating hormone/luteinizing hormone-secreting adenoma,and adrenocorticotrophic hormone-secreting adenoma. The follicle-stimulating hormone/luteinizing hormone adenomas exhibited the lowest STAT3 expression levels.

CONCLUSIONSTAT3 is differentially expressed in pituitary adenoma subtypes, suggesting the cell-specific features of STAT3 regulation,although further investigations are still warranted to clarify the underlying mechanisms.

Adenoma ; Follicle Stimulating Hormone ; Human Growth Hormone ; Humans ; Immunohistochemistry ; Pituitary Neoplasms ; Real-Time Polymerase Chain Reaction ; STAT3 Transcription Factor ; Signal Transduction

OBJECTIVETo compare the efficiency of three different estrogen receptor (ER) immunostaining scoring systems by analyzing the correlation between ER status and clinicopathologic features for prediction of prognosis of patients with endometrial carcinoma (EC).

METHODSER immunostaining (EnVision method) was performed in 160 type I EC and 39 type II EC paraffin samples and was scored by ASCO/CAP criterion, H-Score and Allred scoring system. Correlation between ER status and clinicopathologic features was statistically analyzed.

RESULTSASCO/CAP criterion, H-Score and Allred (cutoff point: 4-8) scoring system showed high concordance in the following aspects. In EC patients, ER status was significantly associated with presurgical CA125 levels (P = 0.015, P = 0.007, P = 0.023), histologic grades (all P < 0.01) and PR status (all P < 0.01). In type I EC cohort, ER status was significantly correlated with PR status (P = 0.008, P < 0.01, P < 0.01) and p53 status (P = 0.042, P = 0.001, P < 0.01). As of the predictive value of ER status for type I EC patient age, ASCO/CAP (P = 0.027) and H-Score criteria (P = 0.035) were both superior to Allred score system (P = 0.064). Among well-known predictive clinicopathologic parameters, including FIGO stage, lympho-vascular involvement, lymph node metastasis, depth of myometrial invasion and omental involvement, ASCO/CAP scoring offered a better correlation (P = 0.005, P = 0.002, P = 0.021, P = 0.067, and P = 0.067, respectively) than H-Score (P > 0.05) and Allred scoring system (P > 0.05).

CONCLUSIONSCompared with H-Score and Allred scoring system, ASCO/CAP criterion is more closely correlated with predictive clinicopathologic parameters. Therefore it may be used as a simple, highly efficient prognostic indicator for EC patients in routine practice.

CA-125 Antigen ; metabolism ; Endometrial Neoplasms ; classification ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; methods ; Lymphatic Metastasis ; Membrane Proteins ; metabolism ; Middle Aged ; Neoplasm Grading ; Neoplasm Invasiveness ; Neoplasm Staging ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Objective To investigate and analyze the effect of Lidocaine Aerosol used in pediatric circumcision.Methods One hundred and twenty cases undergoing selective operation were selected and randomly divided into experimental group and control group with 60 cases in each group.7％ Lidocaine Aerosol local spray and 1％ lidocaine local anesthesia were applied in the experimental group,and 1％ lidocaine local anesthesia only in the control group.And to observe and compare the following contents in the two groups:the fastest heart rate (HR),the lowest oxygen saturation (SpO2),Modified Objective Pain Score (MOPS),visual analogue scale (VAS) and the situation of whether using general anesthesia instead.Results In the experimental group,the fastest HR was (99 ±6.3) beats/min,the lowest SpO2 was (98.4 ± 1.31)％,MOPS score was (2.3 ± 0.39),VAS score was (14.5 ± 4.20),and the control group were of (104 ± 7.7) beats/min,(97.2 ± 1.85) ％,(3.7 ± 0.57),(29.0 ± 5.22),respectively,and these differences were statistically significant between the two groups (t =-6.69,5.58,-4.29,-4.68,respectively; P ＜ 0.05).There were two cases who were given general anesthesia in the experimental group,and eight cases in the control group,and the difference was statistically significant (P ＜0.05).Conclusions The combination of 7％ Lidocaine Aerosol and 1％ lidocaine local anesthesia in the circumcision of children can relieve their pain and stress reaction,which is easily accepted by children patients and their families.

Objective To explore the effect of lidocaine aerosol on alleviating the pain of intravenous puncture before the operation when the local skin spraying anesthesia was used the different time interval and the dose.Methods One hundred and eighty patients undergoing elective operation received the surface spraying anesthesia through the lidocaine aerosol in intravenous puncture site .The patients were divided into A group (4.5 mg) and B (9 mg) group according to the dose of lidocaine aerosol , and the puncture was carried out 3 min, 5 min, 7 min after the medication (A1, A2, A3, B1, B2, B3).The pain was evaluated by numeric pain intensity scale ( NPIS) , and the satisfaction was evaluated by the questionnaire of patients ’ satisfaction in two groups.Results The scores of NPIS in the A and B group were respectively (5.3 ±1.0), (4.9 ±1.1) 3 min after the medication, and the difference was not statistically significant (t=1.17, P>0.05);the scores were respectively (3.9 ±1.2), (2.9 ±1.6) after 5 min, and the difference was statistically significant (t=2.33, P<0.05);the scores were respectively (1.9 ±1.2), (0.9 ±0.7) after 7 min, and the difference was statistically significant (t=2.94, P<0.01).The differences were found in the scores of the NPIS in the A and B group at the different time including 3,5,7 min (F=46.13,56.10, respectively; P<0.01), the degree of pain was relieved in all patients with the extension of time .The rates of satisfaction in the A and B group were respectively 79%, 92%, and the difference was statistically significant (χ2 =6.47, P <0.05 ). Conclusions The effect of lidocaine aerosol on alleviating the pain of intravenous puncture before the operation is obvious.4.5 mg lidocaine aerosol is sprayed twice in surface anesthesia puncture site seven minutes before the injection, which can cause the good analgesic effect .The method of the medication should be reasonably and correctly mastered , which is beneficial to give full play to the analgesia effect and can increase the patients ’ satisfaction of nursing in the operation room .