Objective To evaluate the effects of repeated sevoflurane anesthesia on long-term cognitive function in lactating rats.Methods Twenty-four healthy Spragne-Dawley rats,aged 7 days,weighing 14-17 g,were randomly divided into 3 groups (n =8 each):control group (group C),2.6 ％ sevoflurane group (group S1),and 1.5％ sevoflurane group (group S2).At 7,14 and 21 days after birth,2.6％ and 1.5％ sevoflurane and carrier gas were inhaled for 1 h in groups S1,S2 and C,respectively.Visible plafform trial was carried out on 28 and 29 days after birth,and the swimming speed of the rats was recorded.Place navigation test was performed on 32-36 days after birth,and the escape latency was recorded.Spatial probe test was carried out on 36 days after birth,and the time spent in the platform quadrant,swimming distance and the number of times crossing the platform quadrant was recorded.Results Compared with group C,the escape latency was significantly prolonged in groups S1 and S2 (P ＜ 0.05).The escape latency was significantly longer in group S1 than in group S2 (P ＜0.05).There were no significant differences between the three groups in the swimming speed,time spent in the platform zone,swimming distance and the number of times crossing the platform quadrant (P ＞ 0.05).Conclusion Repeated inhalation of sevoflurane during lactation can impair the long-term declarative memory,which is concentration-related,while it has no effects on the associative learning in rats.

Objective To investigate the relationship of epidermal growth factor receptor (EGFR) mutation with clinical features of baselines as well as serum CEA level in patients with recurrent non-small cell lung cancer (NSCLC). Methods A total of 54 patients with first recurrence of advanced lung cancer who had received chemotherapy were included in this study. ADx-ARMS was performed to detect EGFR gene mutations in surgical specimens taken from the primary tumor. Serum CEA level was measured by the electrochemical luminescence method. Results The mutation rate of EGFR was significantly higher in females than in males (χ2= 11.868, P =0.006), with a total mutation rate of 60.8%in 106 patients. The rate was higher in adenocarcinoma than in other histological types(χ2=6.002,P=0.014), and significantly higher in non-smokers than in smokers (χ2= 8.502,P=0.004) and in the patients with serum CEA level over or equal to 5.0 ng/mL than those with CEA level less than 5.0 ng/mL (χ2=22.543,P=0.000). A multivariate analysis revealed that a higher serum CEA level at the time of disease recurrence was associated with EGFR gene mutations (P = 0.002). Conculsions Serum CEA level is closely associated with the presence of EGFR gene mutations in patients with first recurrence of advanced NSCLC. A higher serum CEA level at the time of disease recurrence is independently associated with EGFR gene mutations. CEA level can be used as a potential indicator to determine EGFR mutation.

Objective To analyze the effect of Noggin silencing on the BMP and Wnt signaling pathways in hair follicle development.Methods The expression of BMP-2, BMP-4, BMPR-IA, BMP-6, BMP-7, LEF-1 andβ-catenin in Noggin silencing MC3T3-E1 stable cell line was detected by RT-PCR and western blot.Results RT-PCR results showed that the expressions of five genes in BMP signaling pathway were all significantly influenced by Noggin silencing, the ex-pressions of BMP-2 (P<0.001), BMP-4 (P<0.01), BMP-6 (P<0.001) and BMP-7 (P<0.001) were all increased and the expression of BMPR-IA (P<0.01) was decreased.While the expressions of the two genes LEF-1 (P<0.001) and β-catenin ( P<0.001) in Wnt signaling pathway were significantly decreased.Western blot results showed that the ex-pressions of these proteins in the two signaling pathways were also affected.The expressions of BMP-2 (P<0.05), BMP-4 (P<0.05), BMP-6 (P<0.05) and BMP-7 (P<0.05) were all increased, while the expressions of BMPR-IA (P<0.05), LEF-1 (P<0.01) andβ-catenin (P<0.001) were decreased.Conclusions There may be a negative feedback regulation of Noggin on the BMP signaling pathway in vitro, but a positive feedback regulation on the Wnt signaling pathway in vitro.It provides certain evidence for studies on the effect of Noggin gene on BMP and Wnt signaling pathways in vivo. There may be an interaction between hair follicle development-related signaling pathways, which still needs further experi-ments to prove.

Objective To explore the effect of doctor-patient interaction based on information platform on hypertensive patients' self-efficacy and treatment compliance in community. Methods A convenience sampling method was used, and 280 patients with hypertension in Huaxin community health service centre were included. Patients were divided into interactive group (89 cases) and control group (191 cases) according to their wishes. Control group received a regular follow-up at community clinics, while interactive group participated in the doctor-patient interaction based on information platform at the same time of regular community clinic visit for 3 months. All patients were investigated using Self-efficacy Assessment Scale for Hypertensives and Treatment Compliance Questionnaire before and after intervention, which was used for effectiveness analysis. Results Before intervention, two groups showed no significant difference in self-efficacy (t=1.635,P>0.05), but there was significant difference in treatment compliance [interactive group:8.00(5.00) vs. control group:10.00(3.00)] (Z=4.409,P<0.05). After the intervention, two groups showed no significant difference in treatment compliance (Z=0.784,P>0.05), but there was significant difference in self-efficacy (interactive group:34.42 ± 4.49 vs. control group:32.63 ± 5.53) (t=2.867, P<0.01), which in interactive group was higher. Before and after the intervention, self-efficacy and the treatment compliance had significantly increased in interactive group (P<0.05). while not significantlyincreased in control group (P>0.05). Before intervention, there was a significant positive correlation between the total score of self-efficacy and treatment compliance(r=0.500, P<0.001), drug therapy compliance (r=0.327, P<0.001) and Non-drug treatment compliance (r=0.469, P<0.001) in two groups, while there was lower positive correlation after intervention. Conclusion The doctor-patient interaction project of community hypertension supervision based on informatization platform is effective on the improvement of self-efficacy and the treatment compliance through the enhanced doctor-patient communication, particularly in promoting the role of self-efficacy to accelerate the improvement of health behaviors, compliance behavior and treatment compliance.

Objective To investigate the role of mitochondrial cytochrome c on hepatocyte apoptosis in non-alcoholic fatty liver disease in rabbits and its pathogenesis.Methods Forty Japanese white rabbits were randomly assigned to control group and model group.The model group was divided into three subgroups: 4-week, 6-week, and 8-week groups, with 10 rabbits in each group.The model groups were subcutaneously injected with peanut oil (1.2 mL/kg), twice a week for 4 weeks, 6 weeks or 8 weeks.The rabbits of all groups were killed at the right time.Serum samples were collected to detect the serum biochemical index levels.Liver tissue samples were taken for pathological observation using HE staining.The hepatocyte apoptosis index ( AI ) was measured by flow cytometry, and mitochondrial permeability transition pore ( MPTP) was evaluated by ultraviolet spectrophotometry.Immunohistochemistry was employed to detect the hepatic expressions of Bcl-2, Bax, CYT C and caspase-3.Western blot was performed to detect the changes of CYT C and caspase-3 protein expressions.Results The model groups showed hepatic injury and high level of TC, TG, CRP, IL-6 and TNF-αbeginning from 4 weeks.With the NAFLD process, the hepatocyte apoptosis index was significantly increased at 4-8 weeks and the MPTP was gradually increased.In the model group, hepatic Bcl-2, Bax, CYT C and caspase-3 expressions were increased steadily with the time passing.Conclusions NAFLD-induced liver damage is associated with apoptosis, and the mitochondrial cytochrome c-mediated apoptotic pathway plays a role in the occurrence of NAFLD.

Objective To study the potential use of the urinary beta-trace protein ( βTP) for diagnosis of type 2 diabetic renal injury.Methods 174 patients with type 2 diabetic mellitus (T2DM) were classified into 2 groups according to the ratio of urinary albumin to creatinine (Alb/Cr):diabetes without renal injury group (group A) and diabetes with renal injury group (group B).70 healthy subjects served as normal control group ( group C).The level of urinary βTP and αl microglobulin (α1MG) was measured by latex particle enhanced immunoturbidimetry assay.The urinary Alb and Cr were determined by nephelometry and Jaffe method respectively.The level of uriuary βTP among all groups was compared and ROC curve analysis was performed.The relevant analysis on urinary βTP,urinary α1MG and other related indexes was made.Results The median level of urinary βTP/Cr in group B was 9.1mg/g Cr,significantly higher than 3.1mg/g Cr of group A and 2.0mg/g Cr of group C.The difference had statistical significance ( H =45.5,P ＜ 0.01).The other indexes ( Alb/Cr,α1MG/Cr,SCr) were all higher in group B than in the other 2 groups ( H =110.9,38.3,11.4 respectively,P ＜0.01).The relevant analysis showed that urinary βTP/Cr was positively correlated with urinary α1MG/Cr (r =0.894,P ＜ 0.01),SCr (r =0.367,P ＜ 0.05 ),HbA(J) C ( r =0.242,P ＜ 0.05 ),systolic pressure ( r =0.162,P ＜0.05 ),and the course of the disease ( r =0.251,P ＜ 0.05 ).No correlation was found between urinary βTP/Cr and diastolic pressure,fasting blood glucose(FBG) or BMI.ROC curve analysis showed the area under the curve (AUC) was 0.86 (95％CI,0.78-0.93)for urinary βTP/Cr and 0.76 (95％ CI,0.67-0.85) for urinary α1MG/Cr.The best cut-off value of urinary βTP/Cr and α1MG/Cr was 4.1mg/g Cr vs 10.9mg/g Cr,the sensitivity was 68.5％ vs 59.7％,and the specificity was 89.8％ vs 80.3％.The difference had statistical significance (P ＜ 0.05).Conclusions Urinary βTP has better diagnostic value for type 2 diabetic patients with renal injury than urinary α1MG.It can sensitively reflect renal tubular injury and can be used as a novel available biomarker to evaluate the renal tubular injury in clinic.

Objective To knockout the murine Txnip gene using microinjection of transcription activator-like effector nuclease ( TALEN) mRNAs.Methods TALEN knockout site recognizing Txnip was designed by tools on line, then constructed the vectors and assayed its cleavage activity at cellular level.TALEN mRNA was transcribed in vitro and microinjected into C57BL/6J mouse zygotes.F0 mice were verified at DNA level with BamHI and TXNIP-knockout mice were obtained.Results We designed and constructed TALENs which recognized and cut the first exon of Txnip, and got four TXNIP knockout mice, among which two were frameshift mutation, demonstrating that the TXNIP-knockout mice were generated by TALEN technique.Conclusions Microinjection of in vitro transcribed TALEN mRNAs into murine zygotes is a highly effective and convenient way to develop TXNIP-knockout mouse model.

Objective To investigate the effect of targeted interruption of cyclooxygenase-2 (COX-2) gene by small interference RNA (siRNA) on the expression of COX-2, vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in eutopic and ectopic endometrial stromal cells (ESC) with endometriosis, and the effect on the content of 6-keto-prostaglandin-F1α (6-keto-PGF1α, metabolites of COX) and the apoptosis of eutopic and ectopic ESC with endometriosis. Methods Ectopic and eutopic ESC from 30 women with endometriosis were isolated and cultured respectively. Then, ESC were classified into three groups: interference group, negative control group and blank control group. ESC in interference group were injected into siRNA transfection complex while ESC in negative control group were injected into negative control transfection complex. ESC from 10 participants without endometriosis were the normal control group. The mRNA and protein expression of COX-2, VEGF, MMP-9 in pre-transfected and post-transfected eutopic and ectopic ESC were detected through real time reverse transcription PCR and western blot. The content of 6-keto-PGF1α was determined by ELISA, the apoptotic cells were detected by flow cytometry. Results After interruption of COX-2 gene, there were no significant difference in the mRNA and protein expression of COX-2, VEGF and MMP-9 between the negative control group and blank control group (P>0.05); the mRNA and protein expression of the three genes in interference group were significantly lower than those in negative control group and blank control group (P<0.05); the mRNA expression of the three genes in interference group of eutopic ESC were 0.87±0.06, 1.76±0.59, 1.04±0.32, in interference group of ectopic ESC were 0.75±0.12, 1.62±0.47, 0.88±0.25, the protein expression of the three genes in interference group of eutopic ESC were 0.457 ± 0.019, 0.500 ± 0.012, 0.361 ± 0.008, in interference group of ectopic ESC were 0.323 ± 0.018, 0.474 ± 0.016, 0.339 ± 0.009;the mRNA and protein expression of the three genes in ectopic ESC had a more reduction than those in eutopic ESC (P<0.05). The results from ELISA revealed that the content of 6-keto-PGF1α in the normal control group [(17.7 ± 1.9) pg/ml] were significantly lower than those in the blank control group (P<0.05), the content of 6-keto-PGF1α in ectopic ESC were significantly higher than that in eutopic ESC (P<0.05), the content of 6-keto-PGF1α in the blank control group of eutopic and ectopic ESC were (32.4±2.6) pg/ml, (38.2±3.7) pg/ml;there was no significant difference in the content of 6-keto-PGF1α between the negative control group and blank control group (P>0.05);compared with those of negative control group and blank control group, the content of 6-keto-PGF1αin interference group decreased significantly (P<0.05), the content of 6-keto-PGF1α in interference group of eutopic and ectopic ESC were (17.1 ± 2.4) pg/ml, (20.9 ± 2.7) pg/ml; the content of 6-keto-PGF1α in eutopic ESC had a slightly more reduction than that in ectopic ESC (P>0.05). The results from flow cytometry displayed that, there was no significant difference in apoptotic cells between the negative control group and blank control group (P>0.05);compared with those of negative control group and blank control group, more apoptotic cells were detected in interference group and the difference was significant (P<0.01);the apoptotic cells in ectopic ESC were significantly more than that in eutopic ESC (P<0.05); the apoptosis rate in interference group of eutopic and ectopic ESC were (33.76 ± 0.06)%, (47.18 ± 0.12)%. Conclusions Our results suggested the targeted interruption of COX-2 gene by siRNA effectively inhibited the mRNA and protein expression of COX-2, VEGF and MMP-9 in both eutopic ESC and ectopic ESC with endometriosis, greatly increased the apoptotic rate of cells and obviously reduced the content of 6-keto-PGF1αby inhibiting the activity of COX-2. And the changes in ectopic endometrium were more evident than those in eutopic endometrium.

Objective To evaluate the effect of docosahexaenoic acid (DHA) on hepatic ischemia-reperfusion (I/R) injury in rats.Methods Fifteen male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were randomly divided into 3 groups (n =5 each) using a random number table: sham operation group (group S), hepatic I/R group (group I/R) , and group DHA.Hepatic I/R was induced by clamping the hepatic pedicle supplying the left and middle lobes of the liver for 60 min, followed by 24 h reperfusion in anesthetized rats.DHA 4 mg/kg was injected intravenously at 30 min before ischemia and 10 min of reperfusion in group DHA.The equal volume of solvent was given instead in S and I/R groups.Blood samples were taken from the inferior vena cava at 24 h of reperfusion for determination of serum alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities, and resolvin D1 concentrations.The rats were then sacrificed, and the livers were removed for determination of myeloperoxidase (MPO) activity (by spectrophotometry), and interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) mRNA expression (by quantitative real-time reverse transcriptase polymerase chain reaction).The livers were cut into sections which were stained with haematoxylin and eosin, and examined under light microscope.Results Compared to group S, the serum ALT and AST activities, serum resolvin D1 concentrations, and MPO activity, and IL-6 and TNF-α mRNA expression in liver tissues were significantly increased in I/R and DHA groups (P＜0.05).Compared to group Ⅰ/R, the serum resolvin 1D1 concentrations, and MPO activity and TNF-α mRNA expression in liver tissues were significantly decreased (P＜0.05) , and no significant difference was found in the serum ALT and AST activities in group DHA (P＞0.05).There was no significant difference in pathological changes of the liver between group DHA and group I/R.Conclusion DHA can attenuate inflammatory responses during hepatic I/R, but it is not sufficient to mitigate liver injury in rats.

Aim To investigate the effect of acacetin on cell proliferation and the influence of acacetin on estrogen receptor expression in vitro.Methods The proliferation rates and the cell cycle changes of acace-tin-treated T47D cells were measured by sulforhodam-ine B(SRB)assay and flow cytometry,respectively. Moreover,the mRNA expressions of estrogen receptor-alpha(ERα),estrogen receptor-beta(ERβ)and pro-liferating antigen(Ki67)were determined by quantita-tive real time PCR (qPCR).Western blot was em-ployed to detect the ERαand ERβprotein expression. Results Acacetin significantly promoted the prolifera-tion and increased the amount of cells arrested in S and G2 /M phase under the concentration of 0.001 ~1 0μmol·L -1 .Ki67 mRNA level and the ERαprotein level in T47D cells were remarkably upregulated after acacetin treatment.To clarify which estrogen receptors played a role in acacetin induced the proliferation of T47D cells,the combination treatment of acacetin and ERαinhibitor (MPP)/ERβ inhibitor (PHTPP) was employed.We found that MPP could reverse the cell proliferation,the cell arrested in S and G2 /M phase and the increased Ki67 mRNA level induced by acace-tin.PHTPP also alleviated the T47D cell proliferation induced by acacetin,whereas no significant changes were found in cell cycle and Ki67 mRNA level.Con-clusion Acacetin stimulates the cell proliferation of T47D cells in the concentration from 0.001 μmol · L -1 to 1 0 μmol·L -1 ,which is mainly mediated by ERα.