OBJECTIVETo construct and analyze the differential expression profiles of microRNAs (miRNA) and mRNA in oral squamous cell carcinoma (OSCC). To explore the miRNA and mRNA of OSCC development and progression.

METHODSThe differential expression profiles of miRNA and mRNA were built by high-throughput deep sequencing technology. Using Gene Ontology (GO) enrichment analysis, the roles of differentially expressed miRNA and mRNA in cell cycle, cell proliferation, cell differentiation and cell apoptosis were analyzed.

RESULTSSeventy-seven differentially expressed miRNA and 1 298 differentially expressed mRNAs were identified in OSCC. GO analysis showed that 73 miRNA had found target mRNA in cell cycle, cell proliferation, cell differentiation and cell apoptosis of OSCC. Moreover, a miRNA could regulate multiple mRNA.

CONCLUSIONThe differential expression profiles of miRNA and mRNA have close relationship with the development and progression of OSCC.

Objective To establish an in vitro accelerated release method of triptorelin acetate microspheres with good in vi-tro/in vivo correlation(IVIVC). Methods The in vivo release of triptorelin acetate from microspheres was obtained by residual method in rats. Influences of pH value,concentration of ethanol,temperature,rotation speed and concentration of antiseptic on the in vitro accel-erated release were studied,then the correlation between in vitro accelerated release and in vivo release of the microspheres was estab-lished by adjusting the release conditions. Results The in vitro accelerated release medium of triptorelin acetate microspheres composed of 15%ethanol solution(containing 0.06%Tween 80 and 0.1%benzalkonium chloride)at 55℃with rotating rate of 200 r/min. The cumulative release of triptorelin acetate from microspheres was 87.35%at 30 h under accelerated release condition,equivalent to in vivo release for 30 days. The established in vitro accelerated release had a good correlation with that of in vivo(y=0.8845x+12.4510, R2=0.9938). Conclusion The in vitro accelerated release of triptorelin acetate microspheres could correlate well with in vivo release and has a potential application in rapid and effective evaluation of triptorelin acetate microspheres.

Objective To investigate the effect of unfractionated heparin (UFH) on the expression of heme oxygenase-1 (HO-1) in intestinal mucosa of mice with sepsis.Methods Thirty-six male C57BL/6J mice were randomly divided into sham group,cecal ligation and puncture (CLP) group and UHF group,n =12 in each group.Model of intestinal injury in sepsis was induced by CLP.In sham group,the mice were exposed without ligation of cecum.In UFH group,the mice were treated intravenously with 8 U of UFH via the tail vein half an hour before the operation and 12 hours after the surgery respectively.Six mice in each group were randomly chosen at 4 hours and 24 hours after operation to collect inferior vena venous blood samples and terminalileum tissues.The serum levels of interleukins (IL-1 β,IL-6),and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA).The serum level of D-lactate was determined by colorimetry.Pathological changes of ileum tissue and Chiu score were observed after hematoxylin eosin (HE) staining.The HO-1 expression was detected immunohistochemically.Results In sham group,no significant changes in the serum levels of IL-1 β,IL-6,TNF-α and D-lactate were observed.Twenty-four hours after the operation,the structure of intestinal mucosa was basically normal without obvious pathology change and no HO-1 positive cells were found.The serum levels of IL-1 β,IL-6,TNF-α,and D-lactate in CLP group were gradually increased,and they were significantly increased as compared with sham group [IL-1 β (ng/L):40.87±2.88 vs.22.60±2.05 at 4 hours,113.73±3.96 vs.22.07±2.74 at 24 hours;IL-6 (ng/L):63.89±3.26 vs.44.89±3.38 at 4 hours,176.56±5.45 vs.45.76±4.02 at 24 hours;TNF-α (ng/L):194.62± 14.13 vs.152.05±6.22 at 4 hours,599.62± 10.20 vs.155.90± 14.18 at 24 hours;D-lactate (mmol/L):0.24± 0.02 vs.0.19 ± 0.01 at 4 hours,0.33 ± 0.04 vs.0.20 ± 0.02 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,edema and inflammation in ileal mucosa,intestinal villi structural damage were observed,the Chiu score was significantly higher than those in the sham group [4.5 (3.0-5.0) vs.0 (0-1.0),P ＜ 0.05],and a small amount of HO-1 positive cells were localized in the intestinal mucosa.Compared with CLP group,the serum levels of IL-1 β,IL-6,TNF-α,and D-lactate of UFH group were significantly decreased [IL-1 β (ng/L):31.53 ± 2.90 vs.40.87 ± 2.88 at 4 hours,61.13 ± 2.80 vs.113.73 ± 3.96 at 24 hours;IL-6 (ng/L):51.16 ± 5.68 vs.63.89 ± 3.26 at 4 hours,81.16 ± 4.54 vs.176.56 ± 5.45 at 24 hours;TNF-α (ng/L):171.76± 5.60 vs.194.62± 14.13 at 4 hours,328.48 ± 10.79 vs.599.62± 10.20 at 24 hours;D-lactate (mmol/L):0.21 ±0.01 vs.0.24±0.02 at 4 hours,0.24±0.02 vs.0.33±0.04 at 24 hours,all P ＜ 0.05].Twenty-four hours after the operation,intestinal injury was ameliorated,the Chiu score was significantly lower [1.5 (1.0-5.0) vs.4.5 (3.0-5.0),P ＜ 0.05],and HO-1 positive cells in the intestinal mucosa was remarkably increased.Conclusion UFH can enhance the expression of HO-1 in intestinal mucosa,reduce the release of inflammatory factors,ameliorate the intestinal inflammatory response,and thus play a protective role in intestinal tissue in mice with sepsis.

Objective To introduce the operation skills of tail vein injection in conscious rats , and improve the success rate of ingection .Methods The rat was fixed by the operator with a self-made binding clothes , one person assists to fix the rat tail, one person performs puncture , and one person performs injection .During the injection process , we should minimize the injury to the rat tail and the stress caused by operation , strictly limit the needle point , the number of puncture , the maximum dose and injection speed , and to make the needle position away from the injection site to avoid contamination and waste of the drug solution .Results The operation method was successfully established and it was fast , stable, with good repeatability and high degree of coordination .Conclusion This operation is rapid, reliable and stable, worthy of recommendation , especially for the intravenous injection of expensive drugs .

Objective:To evaluate the effect of fluoride on the viability of rat ameloblast HAT-7 cells and calcium concentration in the cells.Methods:HAT-7 cells were exposed to NaF at 0,0.4,0.8,1.6,3.2 and 6.4 mmol/L for 24,48 and 72 h respectively. CCK-8 assay was performed to examine the cells proliferation;the apoptosis rate was determined by flow cytometry;Ca2 +concentration in the cells was detected by laser scanning confocal microscopy.Results:The cell proliferation was increased by NaF at 0.4 mmol/L and 0.8 mmol/L,whereas inhibited at 1.6 mmol/L and above.The effects were in a time-dependent manner.NaF increased apoptosis of the cells and increased Ca2 + concentration in the cells in a concentration-dependent manner.Conclusion:Fluoride at low doses promotes proliferation,at high doses inhibits proliferation of HAT-7 cells.NaF of 1.6 mmol/L or more induces apoptosis of HAT-7 cells and in-duce Ca2 + overloading in the cells.

Objective To explore the characteristics of renal cortical perfusion with contrastenhanced ultrasound (CEU)in elderly mice.Methods Randomly selected 12 healthy C57BL/6J young mice(aged 1-2 months)and 12 healthy elderly mice(aged 18-20 months)were assigned respectively into the control group and the experimental group.All mice were examined by color Doppler ultrasound and CEU.Renal morphology and hemodynamics were assessed with conventional ultrasound and CEU was used to assess renal microvascular perfusion,analyzing the time-intensity curve(TIC) and related parameters,including slope rate of ascending curve (A),area under curve (AUC),peak intensity (PI),and time to peak (TTP).Results Conventional ultrasound showed similar blood flow dynamics in the renal hilum artery,the renal segmental artery and the renal interlobar artery between the young and elderly mice groups(all P ＞0.05).Moreover,CEU showed slower ascendance of TIC in the elderly mice group(P ＜ 0.05).Furthermore,the elderly mice were associated with significantly decreased A (0.74 ± 0.06 vs.0.89 ± 0.12,P ＜ 0.05) and increased AUC (1420.08± 208.30)dB · s vs.(1261.41± 34.65)dB/s(P＜0.05),delayed TTP(7.75 ±0.88)s vs.(6.93± 0.44) s (P ＜ 0.05),and similar PI (P ＞ 0.05),compared with those in the young mice group.Conclusions Qualitative analysis of CEU is capable of detecting renal perfusion differences between young and elderly nice,which might lay a basis for further study.

Objective To investigate the clinical value of contrast-enhanced ultrasonography (CEUS)in evaluating renal cortical perfusion in elderly diabetic patients.Methods A retrospective cross-sectional study was conducted.A total of 70 diabetic patients with chronic renal insufficiency stage 1 to 5 were enrolled.The glomerular filtration rate (GFR)was estimated according to the creatinine level.Based on GFR,patients were divided into a mild-to-moderate chronic kidney disease (CKD)group(n=42,CKD stage 1 to 3)and a severe CKD group(n=28,CKD stage 4 to 5).All patients underwent CEUS for the assessment of renal perfusion and renal dynamic imaging for the assessment of GFR.QLAB analysis software was used to obtain the time-intensity curve(TIC)and related parameters including area under the curve(AUC),the ascending slope(A),the derived peak intensity(DP1) and the time to peak (TTP).The differences in renal perfusion parameters were compared between the two groups.Pearson correlation test was used to assess the correlation between the quantitative parameters of renal cortical perfusion and GFR.Results There was no significant difference in age,gender,history of hypertension,basal blood glucose level,systolic or diastolic blood pressure between the two groups (all P ＞ 0.05).CEUS findings revealed high renal cortical blood perfusion in the mild-to-moderate group.There were significant differences in quantitative perfusion parameters including AUC,DPI,A and TTP between the two groups.Pearson correlation test indicated that TTP and A were significantly correlated with GFR in both groups(r =-0.456 and-0.693,0.432 and-0.529,all P＜0.05),while AUC and DPI had no correlation with GFR(r =-0.003 and -0.057,0.066 and-0.081,all P ＞0.05).Conclusions Diabetic patients with mild-to-moderate CKD have high renal cortical blood perfusion.CEUS findings can indirectly reflect renal function.The perfusion parameters A and TTP have good correlations with GFR.CEUS can be used as a safe and effective method to evaluate renal function,especially in patients with severe renal insufficiency,but more studies are needed to verify the results.

Objective To find out parental locus of control and mental health affecting students test anxiety. Methods The samples were from 5 middle schools including 1000 students,and their parents. They were investigated with the general life scales, Sarason's test anxiety scale, Symptom checklist 90 (SCL-90)and Parenting Locus of Control Scale(PLOC). Results The ratio of test anxiety: the mild was 28.8%, moderate was 45.0%, severe was 26.2% ,and felt anxiety was 55.8%. Ratio of test anxiety was higher in the boy students (X2=9.284, P =0.010) ,and heavier(F:16.42±6.70; M:15.18 ±6.51, t=2.716, P=0.007). Student test anxiety was significantly positively correlated with their parental mental health (r fatherl～11=0.166～0.272, Pfather l～11= 0.000;r mother 1～11=0.182～0.242, P mother 1～11= 0.000); Student test anxiety positively correlated with the effectiveness cof parents education, father's belief on the fate, eontrol of fathers for their children, locus of control from fathers (r1～5=0.075～0.143; P1-5=0.000～0.030);felt anxiety positively correlated with self-expect ,pressure from their mothers,over take care attitude from their parents,self-pressure from their parents( r1～6=0.068～0.230; P1-6=0.000～0.050) ,and negatively correlated with respect attitude of parents for their child, Marital relations,Communication time between the students and their parents,attitude of mother for learning( r1-7=-0.074～-0.140;P1～7=0.000～0.034). Felting test anxiety was served as a dependent factor, some factors enter the regression equation,and they were somatization of father, psychotic mother, the pressure from parents, mother over expectations, self-expectations ,the child control from student's father by Logistic regression analysis ( OR1～16=0.675～3.029;P1-6=0.000～0.007). Conclusion Students test anxiety is a common problem in male and female students. Mental health and locus of control from students' parents show test anxiety has somatization of father, psychotic mother, the pressure from parents, mother' expectations,self-expectations, the control of father for the child 6 risk factors.

Objective: To observe the activation of microglia and the changing rule of inflammatory cytokine as IL-6, IL-10 and nuclear factor-κB (NF-κB) in experimental rabbits after spinal cord ischemia reperfusion (SCIR) injury in order to provide theoretical basis for post-conditioning time. Methods: Rabbit SCIR injury model was established by thoracic aorta balloon occlusion. 54 New Zealand male adult white rabbits were divided into 9 groups: Sham group (the animals received balloon implantation without occlusion), SCIR-0h group (reperfusion was conducted at 0 hour of spinal cord ischemia), SCIR-1h, -2h, -3h, -8h, -24h,-48h and -72h groups. n=6 in each group. The number of normal and apoptosis neurons, the levels of Iba-1, IL-6, IL-10 and NF-κB in spinal tissue were examined and compared among different groups respectively. Results: The number of normal neuron was decreasing with the extended reperfusion time, TUNEL-positive neuron began to increasing in SCIR-8h group and the peak was reached in SCIR-24h group. The expression of Iba-1 began to elevating in SCIR-2h group and the peak was obtained in SCIR-8h group; NF-κB began to rising in SCIR-3h group and the peak was observed in SCIR-8h group; both IL-6 and IL-10 arrived the peak in SCIR-24h group. The expressions of NF-κB, IL-6 and IL-10 were positively related to Iba-1 level. Conclusion: Microglia activation had dynamic changes in experimental SCIR rabbits and the expression levels of NF-κB, IL-6 and IL-10 were positively to microglia activation; post-conditioning time at front and back to microglia activation may reduce neuron injury.

Objective: To evaluate the characteristics of renal cortical blood perfusion assessed by contrast-enhanced ultrasound (CEUS) in elderly patients with renal artery stenosis (RAS) and its relationship with renal function. Methods: Ninety-three elderly patients diagnosed with RAS, who were admitted in Beijing Hospital during June 2017 and December 2018, were retrospectively enrolled. According to the degree of RAS, 186 renal arteries were divided into normal renal artery group (n=79), mild RAS group (30% to 49%, n=59), moderate RAS group (50% to 70%, n=33), and severe RAS group (70% to 99%, n=15). Renal cortical blood perfusion and renal glomerular filtration rate (GFR) were measured by CEUS and radionuclide renal dynamic imaging. According to the renal GFR, 186 kidneys were divided into normal renal function group (GFR≥35 ml/min, n=42) and mild renal insufficiency group (35 ml/min>GFR≥25 ml/min, n=51), moderate renal insufficiency group (25 ml/min>GFR≥15 ml/min, n=75) and severe renal insufficiency group (GFR<15 ml/min, n=18). The renal cortical blood perfusion time-intensity curve (TIC) and related parameters were analyzed, including the area under the curve (AUC), the slope of the ascending branch (A), the peak intensity (PI), the peak time (TTP) and the mean transit time (MTT), the kidneys of different RAS groups and patients with different renal function groups were analyzed. Pearson correlation analysis was used to evaluate the correlation between renal cortical blood perfusion parameters and renal GFR. Results: (1) Renal cortical blood perfusion and GFR: CEUS showed that parameter A of TIC was significantly reduced, while TTP was prolonged in the mild renal artery stenosis group compared with the normal renal artery group (both P<0.05), GFP was similar between the two groups. Cortical perfusion parameters, such as AUC, A, PI and GFR were significantly lower, while TTP and MTT were significantly prolonged in the moderate and severe renal artery stenosis group than in the normal and mild stenosis groups (all P<0.05). Compared with the moderate stenosis group, AUC, A, PI and GFR were significantly lower while TTP, MTT were significantly prolonged in the severe renal artery stenosis group (all P<0.05). (2) TIC showed that the renal perfusion parameters, AUC, PI and A were significantly lower, while TTP was significantly longer in the mild renal dysfunction group than in the normal renal function group (all P<0.001). The changes aggravated in proportion with renal dysfunction. (3) Correlation between perfusion parameters and GFR: Pearson correlation analysis showed that the AUC (r=0.774, P<0.05), A (r=0.815, P<0.05) and PI (r=0.772, P<0.05) were positively correlated with GFR; serum creatinine level (r=-0.841, P<0.05), renal function grading (r=-0.731, P<0.05), TTP (r=-0.803, P<0.05) and MTT (r=-0.741, P<0.05) were negative correlated with GFR. The degree of stenosis was negatively correlated with GFR (r=-0.427, P<0.05). Conclusion Cortical perfusion parameters differ significantly among patients with various degree of RAS and renal dysfunction. The renal cortical blood perfusion parameters are correlated with renal GFR.