Objective: To find out the characteristics and regularity trend of influenza activity according to the analyses of influenza surveillance data in Yangzhou from 2012 to 2017, and to provide scientific supports for predicting and controlling the pandemic outbreak of influenza effectively. Methods: The influenza samples were collected from Northern Jiangsu People′s Hospital, Yangzhou First People's Hospital and Gaoyou People’s Hospital, using fluorescent RT-PCR method to detect influenza virus nucleic acid and classifying influenza virus subtypes. Finally, the surveillance data from January, 2012 to December, 2017 of influenza like illness (ILI) cases of Yangzhou were analyzed. Results: Totally 18 083 throat swabs of ILI specimens were collected from 2012—2017 in Yangzhou, 1 983 samples were positive (10.97%), the difference in positive rates of adjacent years was statistically significant (χ²=167.93, P<0.001). In Yangzhou city, influenza virus kept activative in whole year. The influenza epidemic season was winter-spring and summer, accounting for 83.61% of all positive cases, with type A influenza prevalent throughout all year and type B influenza mainly prevalent in spring and winter, each subtype showed alternating prevalence. The influenza virus infection occurred in every age group. The highest positive rate was 16.33% in the age group of 10-19 years and the lowest was 8.52% in the age group of 20-29 years among all detected age groups. Conclusions From 2012 to 2017, the epidemic of influenza in Yangzhou was obviously seasonal and all the subtypes of influenza were in prevalent status alternately. The high-risk groups of Influenza were infants and teenagers who under the age of 19 years. So a long-term influenza surveillance is much needed for the early warning and forecasting the spread of influenza.

Objective To observe the effect of iliopsoas tensile vibration training on the walking ability of stroke survivors.Methods Thirty stroke survivors were randomly divided into an observation group and a control group,each of 15.Both groups were given traditional rehabilitation treatment,while the observation group was additionally provided with iliopsoas tensile vibration training.Both groups were evaluated in terms of the root mean square (RMS) of iliopsoas,active range of motion (A-ROM) of the hip joint,the kinematic parameters of gait and Berg balance scale (BBS) scores before and after the 4-week treatment.Results Before the treatment there were no significant differences between the two groups in any of the measurements.After the treatment improvement in all of the measurements was observed in both groups,with the average RMS iliopsoas,A-ROM,step length,step velocity and BBS score of the observation group significantly better than those of the control group.Conclusions Tensile vibration training of the iliopsoas can significantly improve the muscle excitability of the affected iliopsoas in stroke survivors,and improve their balance and walking ability.

Objective To compare the clinical effect of penehyclidine hydrochloride and atropine in the treatment of acute organophosphorus pesticide poisoning.Methods 86 patients with organophosphorus pesticide poisoning were selected and randomly assigned to the observation group and the control group according to the digital table,43 cases in each group.The two groups were given conventional treatment,while the control group was treated with atropine,the observation group was given penehyclidine hydrochloride.The disappearance time of main symptoms, rescue success rate and adverse reactions of the two groups were observed.Results After salvage treatment, the rescue success rate of the observation group was 97.8%,which of the control group was 88.4%,the difference was statistically significant (x2=1.433,P<0.05).The disappear time of M like symptoms,N like symptoms and central nervous system symptoms in the observation group was significantly shorter compared with the control group (all P<0.05).The number of drugs,dosage,cholinesterase recovery time and hospitalization time between the two groups had statistically significant differences (P<0.05).The incidence rates of blurred vision,restlessness,heart rate and urinary retention in the control group were significantly higher than those in the observation group (all P<0.05).Conclusion Penehyclidine hydrochloride for acute organophosphorus pesticide poisoning can significantly reduce the incidence of symptoms,shorten the disappearance time of symptoms,reduce hospitalization time,improve the efficacy of rescue,reduce the incidence of adverse reactions, it is safe,effective and has great clinical significance.

Objective To investigate the role of Wnt/β-Catenin signaling pathway in the endotoxin induced acute lung injury (ALI) during the treatment by mesenchymal stem cells (MSCs).Methods Six SPF male SD rats were isolated and bone marrow mesenchymal stem cells were cultured.A total of 72 SPF male SD rats with 6-week-old were randomly (random number) divided into 4 groups:control group (n =18) in which phosphate buffered solution (PBS) used instead of lipopolysaccharide (LPS);LPS group (n =18) in which LPS used to induce acute lung injury;LPS + MSCs group (n =18) in which MSCs directly transplanted after injection of LPS;Control + MSCs group (n =18) in which MSCs transplanted after injection of PBS.And then 6 rats of each group were sacrificed at 6 h,24 h,and 48 h separately after injection of LPS.At 24 h after the modeling,lung tissue was taken and the levels of Wrnt signaling pathway components were detected by using immunohistochemistry and Western blot.In addition,quantitative realtime PCR was used to detect the expression of Wnt signaling pathway target genes.Results Compare with the PBS control group,significant decrease in lung dry-to-wet ratio and increase in arterial oxygen partial pressure (PaO2) were found in MSCS transplantation groups.According the immunohistological results,Wnt 5a was significantly increased in the LPS-induced ALI rats and decreased after MSCs transplantation.Moreover,decrease in levels of GSK-3β phosphorylation and β-catenin was found in the lung tissue after MSCs transplantation.In addition,the expressions of Wnt signaling target genes Vegf,Axin2 and Klf4 were decreased significantly after MSCs transplantation.Conclusions In the setting of ALI,the therapeutic effect of MSCs was exerted by decreasing the expressions of Wnt 5a,GSK-3β phosphorylation,β-catenin,and Wnt signaling target genes Vegf,Axin2 and Klf4.Wnt signaling implicated in the therapeutic effect of MSC in the setting of ALI.

OBJECTIVETo investigate the effect of melatonin (MT) on p38 mitogen-activated protein kinase (MAPK) signaling pathway in rats with phosgene-induced lung injury.

METHODSFifty specific pathogen-free male Sprague-Dawley rats were randomly divided into phosgene inhalation group, air control group, saline control group, MT treatment group, and SB203580 (specific inhibitor of p38 MAPK) group, with 10 mice in each group. All groups except the air control group were exposed to phosgene, and the animals were sacrificed 6 h later. Lung wet/dry weight (W/D) ratio and the content of malondialdehyde (MDA) and nitric oxide (NO) and activity of myeloperoxidase (MPO) in bronchoalveolar lavage fluid (BALF) were measured. The qualitative and quantitative expression of p38 MAPK and phospho-p38 MAPK (p-p38) was measured by immunohistochemistry (IHC) and Western blot, respectively. Inducible nitric oxide synthase (iNOS) level in lung tissue was determined by Western blot.

RESULTSCompared with the air control group, the phosgene inhalation group had significantly increased lung W/D ratio and neutrophil count in BALF (P < 0.01); the MT treatment group had significantly lower neutrophil count and lung W/D ratio than the phosgene inhalation group (P < 0.05). IHC demonstrated that the air control group had relatively weak expression of p-p38 in lung tissue; the expression of p-p38 was significantly up-regulated after phosgene inhalation, and it was mainly distributed in infiltrating inflammatory cells and vascular endothelial cells, positive in the cytoplasm and nucleus of many cells. The distribution of p-p38-positive cells in the MT treatment and SB203580 groups was similar to that in the phosgene inhalation group, but the MT treatment and SB203580 groups had a significantly reduced number of cells with p-p38-positive nuclei and a significantly reduced intensity of p-p38 expression signals. The phosgene inhalation group had significantly increased content of MDA and NO and activity of MPO compared with the air control group (P < 0.01); the MT treatment and SB203580 groups had significantly reduced content of MDA and NO and activity of MPO compared with the phosgene inhalation group (P < 0.05), but had higher content of MDA and NO and activity of MPO than the air control group. The Western blot showed that the phosgene inhalation group had significantly increased expression of iNOS and p-p38 compared with the air control group (P < 0.01); the MT treatment and SB203580 groups had lower expression of iNOS and p-p38 than the phosgene inhalation group (P < 0.05).

CONCLUSIONMT and SB203580 have a significant protective effect in rats with phosgene-induced lung injury, and the mechanism may be associated with scavenging free radicals and inhibiting activation of p38 MAPK and expression of iNOS.

Animals ; Bronchoalveolar Lavage Fluid ; Chemical Warfare Agents ; toxicity ; Imidazoles ; Lung ; drug effects ; Lung Injury ; chemically induced ; Male ; Malondialdehyde ; adverse effects ; Melatonin ; physiology ; Mice ; Nitric Oxide ; adverse effects ; Nitric Oxide Synthase Type II ; metabolism ; Phosgene ; toxicity ; Pyridines ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism

ObjectiveTo assess the effects of DNA methyhransferase 1 ( DNMT1 ) gene silencing on DNMTs activity and methylated CpG sites of hMLH-1 in pancreatic cancer cell line PaTu8988.Methods DNMT1 siRNA and negative control siRNA was constructed by Ambion Company of United States.Then they were transfected into pancreatic cancer cell line PaTu8988 at the concentrations of 15,30 nmol/L,and the cells without transfection was used as the control group.Real-time PCR and Western blotting were applied to detect the DNMT1 mRNA and protein expression,and DNMTs activity was detected by using DNMTs activity assay kit.Change of methylation of CpG island of hMLH-1 was detected by bisulfite sequencing PCR (BSP).The expression of hMLH-1 mRNA was detected by Real-time PCR.ResultsAt 48 h after transfection,Realtime RT-PCR analysis showed that the levels of DNMT1 mRNA in DNMT1 siRNA group ( 15 nmol/L) and DNMT 1 siRNA group (30 nmol/L) were 0.573 ± 0.026 and 0.143 ± 0.044,which were significantly lower than those in control group 1.020 ±0.217 and negative siRNA 15 nmol/L group 0.900 ±0.475,and negative siRNA 30 nmol/L group 0.938 ± 0.327 (P ＜0.05 ).Western blotting analysis showed that the level of DNMT1 protein of DNMT1 siRNA group was also lower than those of negative siRNA and control groups.DNMT activity in DNMT1 siRNA15,30 nmol/L groups was 0.364 ± 0.124and 0.250 ± 0.072,which were significantly lower than those in control group 0.931 ± 0.065and negative siRNA group 0.665 ± 0.055 and 0.472 ± 0.040.DNMT activity was positively correlated with DNMT1 mRNA expression ( r =0.69,P ＜ 0.01 ).DNMT1 RNA interference decreased 8 methylated CpG sites of hMLH-1 to 1 site.Concluslons DNMT1siRNA can specifically inhibit the expression of DNMT1 gene of PaTu8988 and DNMT activity,and can decrease methylated CpG sites of hMLH-1 gene.

Objective To investigate the effects of histone deacetylase 1 (HDAC1) gene silencing on cell cycle of pancreatic cancer cell line PaTu8988 and possible mechanism.Methods The PaTu8988 cells were routinely cultured and divided into control group,negative control siRNA group (c-siRNA),HDAC1 siRNA 15 nmol/L group and HDAC1 siRNA 30nmol/L group.After Liposomes 2000 transfection for 48 h,the Western blotting was used to detect the efficiency of HDAC1 gene silencing on protein levels and the expression of p21 protein.Cell cycle was evaluated by using flow cytometry.Results Compared with that of control group,the expression of HDAC1 protein in PaTu8988 cell of HDAC1 siRNA 30nmol/L group was significantly decreased,and the expression of p21 protein was significantly increased; the percentage of G2/M phase cells were significantly decreased[(21.48 ±3.67)％ vs.(28.28 ±2.94) ％,P＜0.05]; while the percentage of S phase cells were significantly increased[( 50.20 ± 6.85 ) ％ vs.( 32.49 ± 2.78 ) ％,P ＜ 0.05].Conclusions HDAC1 siRNA can efficiently and specifically inhibit the expression of HDAC1 in PaTu8988 cells,and can induce S phase cells arrest,which may be related with up-regulation of p21 protein expression.

Objective To investigate the expressions of DNA methyltransferase 1 (DNMT1) in pancreatic carcinoma and its clinical significance.Methods 30 samples of pancreatic cancer tissues and paired para-cancerous tissues were collected from patients who underwent curative pancreatectomy.The levels of DNMT1 mRNA were detected by real-time RT-PCR.Expressions of DNMT1 protein were detected by streptavidin peroxidase immunohistochemistry.The relationships between expression of DNMT1 and clinicopathological findings were analyzed.Results The value of relative quantification (RQ) of DNMT1 mRNA in human pancreatic cancer tissues was 2.32 (1.17 ～ 5.17 ), which was significant higher than 0.78 (0.07 ～3.14) in para-cancerous tissues(P ＜0.05).The index of expression of DNMT1 protein in human pancreatic cancer tissues was (54.5 ±21.2)% ,which was significant higher than( 10.9 ± 15.0)% in paracancerous tissues (P ＜ 0.01 ).Patients were divided into the high DNMT1 group (n = 19) with above 54.5% of the DNMT1 positively cancer cells and the low DNMT1 group ( n = 11 ) with less than 54.5% of the DNMT1 positively cancer cells.The high expression of DNMT1 was correlated significantly with clinical staging ( x2 =6.897, P = 0.029), lymph node metastasis ( x2 = 4.739, P = 0.029) and neural invasion ( x2 = 5.44, P =0.020).On the other hand, no association between DNMT1 expression and age, gender, tumor location,tumor size, tumor differentiation, the serum CEA and CA19-9 levels could be found.Conclusions DNMT1 mRNA and protein was highly expressed in pancreatic cancer tissues.High expression of DNMT1 might be related to the aggressiveness of pancreatic cancer, lymph node metastasis and neural invasion.

Objective To investigate effects of histone deacetylase 1 (HDAC1) gene silencing on proliferation and apoptosis of pancreatic cancer cell line PaTu8988. Methods The PaTu8988 cells were cultured and divided into control group (untreated), negative control group (treated with 30 nmol/L negative siRNA), low HDAC1 group (treated with 15 nmol/L HDAC1 siRNA) and high HDAC1 group (treated with 30 nmol/L HDAC1 siRNA). The real-time PCR and Western blot were used to detect the efficiency of HDAC1 gene silencing on mRNA and protein levels ,respectively, at 48 hours after transfection of HDAC1 siRNA. Cell proliferation and apoptosis were evaluated using cell counting kit and flow cytometry, respectively. Results Forty-eight hours after transfection of HDAC1 siRNA, the expression of HDAC1 mRNA level in PaTu8988 cells was 46.1%±6.1% in low HDAC1 group and 32.3%±1.4% in high HDACI group, which were lower than that in control group (100.0%±3.4%) and negative control group (87.4%±28.3%,P<0.05). The expression of HDAC1 protein was higher in control and negative control groups than in low and high HDAC1 groups. Cell survival rate was 100.0%±17.1% in control group, 87.1%±5.0% in negative control group, 68.7%±4.7% in low HDAC1 group and 61.6%±2.0% in high HDAC1 group with significant difference (P<0.05). Meanwhile, the cell apoptic rate in control (4.20%±0.95%) and negative control (4.59%±1.26%) groups was lower than that in low (10.09%±1.36%) and high (11.19%±6.07%) HDACI groups (P<0.05). Conclusions HDACI siRNA can effectively and specifically inhibit the expression of HDAC1 and proliferation of PaTu8988 cells and induce cell apoptosis.

Objective To investigate the effects of trichostatin A (TSA) on cell proliferation and cell cycle in human gastric cancer cell line SGC-7901 in vitro and its mechanism. Methods SGC-7901 cells were treated with 0.1, 0.5 and 2.0 μmol/L of TSA for 24 hrs. Growth inhibition rates of cells were measured by MTT assay and cell cycles were detected by flow cytometery (FCM). Expressions of cyclin D1 and p21 mRNA were measured by real-time PCR. Results The proliferations of SGC-7901 cells were inhibited when treated with TSA for 24 hrs. The inhibition rates in groups treat with 0.1, 0.5 or 2.0 μmol/L of TSA were 3.52%±6.11%, 13.29%±4.13% or 14.24%±2.80% ,respectively. The cell percentage of G0/G1 phase were higher in 0. 5 pznol/L group (71.26%±0.51%) and 2.0 μmol/L group (71.03%±0.12%) compared with control group (51.12%±1.17%). The cell percentage of S phase were lower in 0.5 μmol/L group (13.55%±0.44%) and 2.0 μmol/L group (10.63%±0.63%) compared with control group (34.60%±0.60%). The expression of cyclin D1 mRNA was down-regulated, whereas p21 mRNA expression was up-regulated. Conclusions TSA inhibits SCG-7901 gastric cancer cell proliferation by affecting the cell cycle control gene eyclin D1 and p21 mRNA expressions, which induce G0/G1 cell phase cycle arrest and ultimately impact on the growth of tumor cells.