OBJECTIVE To explore the effect of clofenotane (DDT) on epithelial-mesenchymal transition (EMT) and the relevant molecular mechanism in human colorectal cancer cells. METHODS Human colorectal cancer cells DLD1 were treated with DDT 0.01, 0.1, 1.0, 10.0 and 100.0 nmol·L-1 for 48 h. Then, the morphology of DLD1 cells was observed. mRNA levels of E-cadherin, N-cadherin, vimentin and Snail1 were detected by real-time PCR. Protein expression of STAT3 signaling pathway of proteins STAT3 and p-STAT3 was detected by Western blotting. STAT3 inhibitor WP1006 (5μmol · L-1) was added to determine its impact on DDT-induced alternation of STAT3/Snail1 signaling and EMT-related molecules. Protein expression of STAT3 and p-STAT3 was detected by Western blotting and mRNA levels of E-cadherin, N-cadherin, Vimentin and Snail1 were detected by real-time PCR. RESULTS DLD1 cell morphology was changed after exposure to DDT 0.01-100.0 nmol · L- 1. Meanwhile, real-time PCR showed that the mRNA level of E-cadherin was significantly decreased compared with normal cell control (P<0.01), which was 42.4±2.8%of that in the normal control group. The mRNA levels of N-cadherin, Vimentin and Snail1 were significantly increased (P<0.01), which were 1.91±0.1, 1.5±0.2 and 1.5±0.1 times that of the normal control group. DDT 0.1, 1.0 and 10.0 nmol · L-1 exposure induced up-regulation of STAT3 and p-STAT3 protein levels (P<0.01), which were 2.1 and 1.8 times that of the normal control group. The addition of STAT3 inhibitor WP1066 (5 μmol · L-1) prevented STAT3 from phosphorylation as well as the up-regulation of Snail1(P<0.01), which was (56.3 ± 0.9)% that of the DDT 1.0 nmol · L-1 treat?ment group. Compared with DDT treatment alone, the mRNA levels of EMT-related molecules were remarkably reversed by WP1066 (5 μmol · L- 1) co-treatment, increasing E-cadherin but decreasing N-cadherin and vimentin in DLD1 cells(P<0.01), which were 50.2±2.9%and 61.6±6.1%of those in the DDT 1.0 nmol · L- 1 treatment group, respectively. CONCLUSION DDT alters the expressions of EMT-related molecules including E-cadherin, N-cadherin and vimentin via STAT3/Snail1 signaling, thus promoting the EMT process in human colorectal cancer cells. This progress may be closely related to DDT-induced colorectal cancer development.

Objective To compare the adverse reactions of intranasal and intravenous dexmedetomidine on tracheal extubation during wake up of general anesthesia.Methods One hundred and twenty patients who ASA Ⅰ or Ⅱ grade were divided into four groups (each 30 patients) by random digits table method.The patients in intravenous group were given 0.5 μ g/kg intravenous dexmedetomidine (diluted to 10 ml by 0.9％ sodium chloride,intravenous injection slowly,≥30 s).The patients in intranasal group 1 were given 0.5 μg/kg intranasal dexmedetomidine.The patients in intranasal group 2 were given 0.8 μg/kg intranasal dexmedetomidine.The patients in control group were given intravenous 0.9％ sodium chloride.The systolic blood pressure(SBP),mean arterial blood pressure (MAP),heart rate were compared among groups.Eyes open time and extubation time,the rate of cough and the degree during extubation were compared too.Results The SBP,MAP,heart rate in intravenous group,intranasal group 1 were significantly higher than those in basal state (P ＜ 0.05).The SBP,MAP,heart rate at different time in intranasal group 2 had no significant difference (P ＞ 0.05).The SBP,MAP,heart rate before extubation and after extubation for 3 min in control group were significantly higher than those in intravenous group,intranasal group 1 and intranasal group 2 (P ＜ 0.05).Eyes open time and extubation time among four groups had no significant difference(P ＞0.05).The rate of cough,restlessness and 3 scores of degree before extubation in intravenous group,intranasal group 1 and intranasal group 2 were significandy lower than those in control group [43％ (13/30),50％(15/30),47％(14/30) vs.70％ (21/30); 17％(5/30),23％(7/30),20％(6/30) vs.43％(13/30);53％ (16/30),60％ (18/30),50％ (15/30) vs.80％ (24/30)] (P ＜ 0.05).Conclusions Either intravenous or intranasal dexmedetomidine can effectively prevent the stress reaction during extubation,decrease the degree of restlessness and cough.Intranasal dexmedetomidine(0.8 μ g/kg) is more effective and safe.

In this study, real-time PCR and immunohistochemistry were used to detect coxsakie and adenovirus receptor (CAR) expression. Both localization and quantity were evaluated in the uteri obtained at days post coitus (dpc) 2.5, 4.5, 6.5, 8.5. Outcome of PCR was assessed by 2(-ΔΔCt) method. Image Pro-Plus 6.0 software was used for quantifying mean density of CAR expression in immunohistochemical sections. We found relatively weak CAR expression in the mouse uteri during implantation window. PCR and immunohistochemistry revealed highest CAR expression was detected on dpc 2.5 followed by down-regulation of CAR at dpc 4.5 and 6.5 (with significant difference). At dpc 8.5, CAR expression was increased slightly again. It is concluded that during implantation, the expression of CAR mRNA and protein is declined, resulting in the impairment of tight junction between cavity epithelium cells. After implantation window closure, CAR appears again to maintain epithelium stability. CAR might play an important role during embryo implantation procedure.

Blastocystis sp. is a kind of protozoa living in the intestinal tract of human and animals, which will cause intestinal diseases such as diarrhea, abdominal distension and vomiting. This paper was aimed to understand the infection of Blastocystis sp. In golden monkeys and the transmission path in North China. Thirty-seven feces samples from golden monkeys and 116 cockroach samples from Shijiazhuang Zoo were collected from July to October 2019 for PCR analysis of Blastocystis sp. Genetic diversity analysis was further conducted on the samples with positive PCR results. The results showed that the infection rate was 48.7% (18/37) in golden monkeys and 82.8% (96/116) in cockroaches, respectively. The genetic evolution analysis based on small subunit ribosomal RNA demonstrated that three subtypes (ST) of Blastocystis sp. including ST1, ST2, and ST3 existed in the intestinal tract of golden monkeys, while only ST2 was detected in the intestinal tract of cockroaches. This paper may provide supports for the quarantine and control of Blastocystis sp. for the zoo in Northern China.

Objective To analyze the expression levels and clinical significance of N-myc down-stream-regulated gene 3(NDRG3)and semaphoring 3A(SEMA3A)in elderly patients with acute ischemic stroke(AIS).Methods A total of 100 elderly AIS patients admitted to Department of Geriatrics of Liaocheng People's Hospital from September 2020 to September 2022 were included as the study group.According to their NIHSS score at admission,they were divided into mild(34 cases),moderate(31 cases)and severe(35 cases)subgroups.All patients were followed up for 3 months after discharge.And they were assigned into good prognosis group(69 cases)and poor prognosis group(31 cases)based on the modified Rankin scale score.Another 100 healthy individ-uals who underwent physical examination in our hospital during the same period were recruited as the control group.Western blotting was used to detect the protein expression of NDRG3 and SEMA3A in peripheral blood mononuclear cells(PBMC).ELISA was applied to measure the con-tents of VEGF,TGF-1,TNF-α,and IL-17 in peripheral blood samples.Spearman rank correlation analysis was performed to analyze the correlation of NDRG3 and SEMA3A levels with NIHSS score,and ROC curve was plotted to analyze the values of NDRG3 and SEMA3A in predicting poor prognosis in elderly AIS patients.Results The expression levels of NDRG3 and SEMA3A in PBMC were obviously higher in the study group than the control group(1.11±0.16 vs 0.76± 0.13,0.78±0.13 vs 0.42±0.09,P＜0.01).The levels in the mild,moderate and severe subgroups were significantly higher than that of the control group(P＜0.01).The poor prognosis group had statistically higher expression levels of NDRG3 and SEMA3A than the good prognosis group(P＜0.01).Spearman rank correlation analysis showed that the NIHSS score was positively corre-lated with the expression levels of NDRG3 and SEMA3A in elderly AIS patients(r=0.597,P＜0.01;r=0.618,P＜0.01),while the NDRG3 level was positively correlated with that of SEMA3 A(r=0.477,P＜0.01).ROC curve analysis indicated that the AUC value of combined NDRG3 and SEMA3A levels was superior to that of NDRG3 and SEMA3A alone in predicting of poor progno-sis(0.962 vs 0.861,0.880,P＜0.01).Conclusion The levels of NDRG3 and SEMA3A proteins are up-regulated in elderly AIS patients,and are closely associated with the severity and prognosis of the disease.

Objective: To observe the expression level of microRNA-181 (miR-181) and importin-α3 in oxidized low density lipoprotein (ox-LDL) induced vascular endothelial cell injury models, and explore the effect and mechanism of miR-181 on endothelial cell injury. Methods: Human vein endothelial cell line CRL-1730 were cultured and vascular endothelial cell injury model was established by intervention with ox-LDL. The cells were divided into control group (intervened by double distilled water), low-dose group (intervened by 10 μg/ml ox-LDL) and high-dose group (intervened by 20 μg/ml ox-LDL). In addition, cells of low-dose group were divided into miR-181 mimic group (miR-181 mimic was transfected) and mimic control group (miR-181 mimic control was transfected). Cell viabilities, mRNA and protein expression level of interleukin-6 (IL-6), miR-181, importin-α3, and nuclear transcription factor-κB (NF-κB) were measured by methyl thiazolyl tetrazolium (MTT), real-time PCR and Western blot, respectively. Results: (1) The cell viabilities in low-dose group and high-dose group were lower than control group (0.207±0.012 and 0.204±0.007 vs. 0.323±0.018, all P<0.01). The relative IL-6 mRNA expression in low-dose group and high-dose group were higher than control group (1.24±0.16 and 1.36±0.23 vs. 0.22±0.03, all P<0.01). The relative miR-181 mRNA expression in low-dose group and high-dose group were lower than control group (0.91±0.11 and 0.88±0.07 vs. 2.20±0.13, all P<0.01). The relative importin-α3 mRNA expression in low-dose group and high-dose group were higher than control group (1.23±0.22 and 0.55±0.03 vs. 0.44±0.06, all P<0.01). The relative NF-κB mRNA expression in low-dose group and high-dose group were higher than control group (1.67±0.34 and 0.41±0.11 vs. 0.11±0.04, all P<0.01). The relative importin-α3 protein expression in low-dose group and high-dose group were higher than control group (1.44±0.23 and 1.31±0.22 vs. 0.29±0.08, all P<0.01). The relative NF-κB protein expression in low-dose group and high-dose group were higher than control group (0.43±0.05 and 0.37±0.04 vs. 0.16±0.03, all P<0.01). (2)The cell viabilities in miR-181 mimic group was higher than in mimic control group (0.262±0.008 vs. 0.211±0.021, P<0.01). The relative miR-181 mRNA expression level in miR-181 mimic group was higher than in mimic control group (4.23±0.34 vs. 0.88±0.16, P<0.01). The relative importin-α3 mRNA expression level in miR-181 mimic group was lower than in mimic control group (0.24±0.03 vs. 1.08±0.13, P<0.01). The relative NF-κB mRNA expression level was lower in miR-181 mimic group than in mimic control group (0.13±0.03 vs. 0.51±0.06, P<0.01). The relative importin-α3 protein expression level was lower in miR-181 mimic group than in mimic control group (0.34±0.06 vs. 1.67±0.26, P<0.01). The relative NF-κB protein expression level was lower in miR-181 mimic group than in mimic control group (0.43±0.02 vs. 1.53±0.36, P<0.01). Conclusions Lower miR-181 expression but higher importin-α3 and its downstream NF-κB signaling are associated with ox-LDL induced vascular endothelial cell injury and up-regulation of miR-181 could alleviate ox-LDL induced vascular endothelial cell injury possibly via importin-α3/NF-κB pathway.

Objective To clarify the necessity of therapeutic drug monitoring (TDM ) of voriconazole ,and give relevant clinical tips ,by comparing the plasma concentration of different clinical specialties before and after adjustment of dose .Methods This is a retrospective study of voriconazole TDM data .It involves 435 cases voriconazole plasma trough concentration meas-urement results of 154 inpatients to make a preliminary assessment .Results 4 .3% plasma concentration were higher than 5 .5 μg/ml ,26 .5% plasma concentration were less than 1 .0 μg/ml in renal transplantation department ;while 52 .3% plasma concentration were higher than 5 .5 μg/ml ,no less than 1 .0 μg/ml in infectious disease department .Conclusions Therapeutic drug monitoring is necessary for rational use of voriconazole .The majority of plasma concentrations in renal transplantation pa-tients were <1 .0 μg/ml ,lower than recommended treatment concentration range ;while most infectious disease patients have> 5 .5 μg/ml ,higher than recommended treatment concentration range .Clinical pharmacists can be more closely involved in the clinical use of voriconazole based on the results of the therapeutic drug monitoring .

In this study,real-time PCR and immunohistochemistry were used to detect coxsakie and adenovirus receptor (CAR) expression.Both localization and quantity were evaluated in the uteri obtained at days post coitus (dpc) 2.5,4.5,6.5,8.5.Outcome of PCR was assessed by 2-△△Ct method.Image Pro-Plus 6.0 software was used for quantifying mean density of CAR expression in immunohistochemical sections.We found relatively weak CAR expression in the mouse uteri during implantation window.PCR and immunohistochemistry revealed highest CAR expression was detected on dpc 2.5 followed by down-regulation of CAR at dpc 4.5 and 6.5 (with significant difference).At dpc 8.5,CAR expression was increased slightly again.It is concluded that during implantation,the expression of CAR mRNA and protein is declined,resulting in the impairment of tight junction between cavity epithelium cells.After implantation window closure,CAR appears again to maintain epithelium stability.CAR might play an important role during embryo implantation procedure.

Objective:To explore the vasculopathy of central nervous system (CNS) infections in critically ill children.Methods:The clinical data of 43 children with CNS infections requiring mechanical ventilation admitted to the Intensive Care Unit of Beijing Children′s Hospital from July 2017 to October 2018 were analyzed retrospectively.The peak systolic velocity and the pulsatility index of the bilateral middle cerebral artery in each child were detected by Transcranial Doppler (TCD), and the vasculopathy was explored by combining with the imaging manifestations such as computed tomography or magnetic resonance imaging.Results:TCD results were normal in 11 cases and abnormal in 32 cases.In the abnormal group, the cerebral blood flow velocity increased in 27 cases and decreased in 5 cases.Of the 27 cases with increased velocity, 15 cases met the criteria of vasospasm, and 4 of the 5 cases with decreased velocity showed shock waves.Among all patients, 21 cases (48.9%) had cerebral hypoxic-ischemic manifestations, 6 cases (13.9%) had cerebral hemorrhage (5 cases were accompanied with cerebral ischemia), and 2 cases (4.7%) were complicated with cerebral malacia during hospitalization.Twenty cases in all of them had completed magnetic resonance angiography, which was abnormal in 14 cases (70%). There were 32 cases (74.4%) with poor prognosis at discharge, including 10 cases of death or brain death (23.3%, 10/32 cases).Conclusions:Cerebral hemodynamic abnormalities and ischemic changes are more common in children with severe encephalitis.Patients with vasospasm or decreased flow velocity have the worst prognosis.

Objective:To investigate the predictive value of abnormal heart rate circadian rhythm for all-cause mortality in stage 5 chronic kidney disease (CKD 5) patients.Methods:The retrospective study was performed in CKD 5 patients enrolled from the First Affiliated Hospital of Nanjing Medical University (Jiangsu Province Hospital) and the Affiliated BenQ Hospital of Nanjing Medical University from February, 2011 to December, 2019. A total of 159 healthy volunteers were enrolled as the healthy control group during the same period. The circadian rhythm of heart rate was monitored by 24-hour Holter. Related indices (including 24-hour, daytime and nighttime mean heart rate, night/day heart rate ratio, 24-hour maximum heart rate, 24-hour minimum heart rate and difference between maximum and minimum of 24-hour heart rate) were calculated. Non-dipping heart rate was defined as night/day heart rate ratio greater than 0.9. Cox regression model was used to analyze the risk factors of all-cause mortality in CKD 5 patients. Kaplan-Meier survival curve and Log-rank test were used to compare the differences of cumulative mortality between high ratio group (night/day heart rate ratio>0.91) and low ratio group (night/day heart rate ratio≤0.91). The nonlinear relationship between night/day heart rate ratio and all-cause mortality was analyzed by restricted cubic spline plot. Time-dependent receiver operating characteristic (ROC) curve was used to analyze the predictive value of night/day heart rate ratio for all-cause mortality in CKD 5 patients.Results:A total of 159 healthy volunteers and 221 CKD 5 patients were included in this study. There were 123 males (55.66%) and the age was (52.72±13.13) years old in CKD 5 patients. The total median follow-up time was 50.0 months. Compared with controls, 24-hour, nighttime mean heart rate, 24-hour minimum heart rate in CKD 5 patients were increased (all P<0.05), furthermore, the night/day heart rate ratio was higher [(0.91±0.09) vs (0.81±0.08), P<0.001], showing "non-dipping heart rate". However, the 24-hour maximum heart rate and the difference between maximum and minimum of 24-hour heart rate in CKD 5 patients were lower than controls (both P<0.05). Multivariate Cox regression analysis showed that the increased night/day heart rate ratio (per 0.1 increase, HR=1.557, 95% CI 1.073-2.258, P=0.020) was an independent influencing factor for all-cause mortality in CKD 5 patients. Kaplan-Meier survival curve analysis showed that the cumulative mortality of the high ratio group was significantly increased than that of the low ratio group (Log-rank test χ 2=7.232, P=0.007). From the restricted cubic spline plot, there was a linear effect between night/day heart rate ratio and all-cause mortality ( P=0.141), and when night/day heart rate ratio was above 0.91, the risk of all-cause mortality was significantly increased in CKD 5 patients. According to time-dependent ROC curve, the accuracy of night/day heart rate ratio in predicting all-cause mortality was 70.90% even when the survival time was up to 70.0 months. Conclusions:The circadian rhythm of heart rate in CKD 5 patients displays "non-dipping" state. High night/day heart rate ratio is an independent influencing factor for all-cause mortality in CKD 5 patients.