Objective To investigate the effects of azithromycin on hyperoxia induced lung injury in neonatal rats.Methods Ninety 2-day-old neonatal rats were randomly divided into 3 groups (30 cases in each group):air control group,hyperoxia model group,azithromycin treated group.The rats in air control group were exposed to indoor air,and the rats in hyperoxia model group and azithromycin group were exposed to 900 mL?L-1 O2 for 14 days.From 2-days-after birth,rats in treated group received intraperitoneal injection of azithromycin (200 mg?kg-1)per day,rats in air control group and hyperoxia model group received intraperitoneal injection of normal saline.Ten rats in each group were executed and their lung tissues were taken out and were observed under a optical microscope on the 3rd,7th,14th days from the start.The expression of interferon gamma(IFN-?),connective tissue growth factor(CTGF) and matrix metalloproteinase-9 (MMP-9) in the lung tissues were detected by immunohistochemistry.Results There was no expression of IFN-? positive cells in air control group,but the expressional intensity level was gradually enhanced as the time of inhaling hyperoxia prolonged in hyperoxia model group,it reached the peak on the 7th day,and then obviously declined on the 14th day,but still higher than that of air control group.The expressional intensity level of IFN-? in azithromycin treated group was lower than that of hyperoxia model group in each time segment,there were significant differences among them(Pa

China ; epidemiology ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Humans ; Listeria monocytogenes ; physiology ; Listeriosis ; epidemiology ; Serotyping

Objective To discuss the clinical effect of somatostatin combined with psychological intervention in the treatment of severe acute pancreatitis. Methods 48 cases of severe acute pancreatitis patients in Zhejiang Traditional Chinese Medical hospital from January 2012 to December 2016 were divided into the experimental group 24 cases and the control group 24 cases, the control group was given routine clinical treatment, the experimental group on the basis of the control group was given psychological intervention and somatostatin, the treatment effect of the patients between two groups were compared and analyzed. Results APACHEⅡ score, decline degree of Serum amylase, mortality, hospitalization time, remission time and complication rate in the experimental group was significantly better than the control group, the difference between groups was statistically significant (P<0.05). Conclusion In the treatment of severe acute pancreatitis, the combination of somatostatin and psychological intervention could effectively alleviate the clinical symptoms and control the development of the disease.

PBL teaching method is a new mode of teaching which is originated from the West and implemented into China in recent years with an expectation that it would mainly develop the students’ self-learning ability,and enhance their skills of comprehensive thinking and solving actual problems.The author summarizes the practical experience of using PBL teaching methods in the theory teaching in Department of Medical Microbiology and Human Parasitology,China Medical University in the past three years,and then proved this method is very helpful to improving the students’integrated thinking by analysis of sample.At the same time the results also suggested that the students showed high enthusiasm in discussing the cases.By this way,the students showed great subjective intiative in their studies.

Abstract: Objective　To analyze the epidemic characteristics of hand, foot and mouth disease (HFMD) in Nanping City, Fujian province and to provide the basis for formulating effective prevention and control measures as well as evaluating the efficacy of prevention and treatment. Methods　Descriptive epidemiological method was used to analyze the incidence data of HFMD in Nanping City from 2012 to 2021. Results　A total of 49 231 cases of HFMD were reported in Nanping City from 2012 to 2021. The incidence fluctuated greatly over the 10-year period, ranging from 76.10/100 000 to 308.93/100 000, with an average incidence of 184.99/100 000 per year. The overall incidence and the number of cases showed a fluctuating downward trend over time, but the incidence was high in the next year, and there were statistically significant differences in the incidence rates between different years(χ2=8 169.176, P<0.001). There were significant regional differences in the incidence, the top three average annual incidence rates were: Guanze County (370.76/100 000), Zhenghe County (295.31/100 000) and Wuyishan City (250.31/100 000). There were two peaks of HFMD incidence each year, with the first occurring in May and June and the second occurring in September and October. The incidence rate was higher among males (215.86/100 000) than females (152.93/100 000), and males were more susceptible than females (RR=1.412, 95%CI=1.387-1.438). The cases were mainly aged 0-4 years, accounting for 86.25% (42 461/49 231) of all cases, and the incidence rate gradually decreased with increasing age (χ2trend=570,105.801, P<0.001). The majority of cases (85.22%, 41 953/49 231) occurred in children living in scattered areas, followed by children in kindergartens (12.39%, 6 101/49 231). The etiological results showed a total of 3 476 laboratory-confirmed cases, and the proportion of three (classes) of enterovirus positivity varied each year, with different pathogen compositions showing statistical significance (χ2=584.613, P<0.001). In addition to the years 2015-2017, during which Cox A16 and EV71 were the dominant strains, other years were dominated by other enteroviruses, with EV71 being the main type in severe and fatal cases of HFMD in Nanping City. Conclusion　Nanping City should strengthen health education for children living in the diaspora and in day-care centers, enhance surveillance of epidemics and pathogenology, improve vaccination rates against EV71, focus on the detection and typing of other enteroviruses, and implement effective prevention and control measures for HFMD.

ORF1 and ORF2 gene of porcine circovirus type 2 were cloned by PCR with the specific primers designed according to genome of PCV2 (AY035820). Following extraction and digestion, PCR products were subsequently inserted into universal transfer vector plECMV (deleted partial gE and gI of pseudorabies virus) to generate recombinant transfer plasmid pIEORF1-ORF2. The genomic DNA of PRV TK-/gE- /LacZ+ strain and pIEORF1-ORF2 were co-transfected into IBRS-2 cells with lipofectin, and recombinant virus TK- /gE- /gI- /ORF1-ORF2+ was selected by PCR with ORF1 gene and ORF2 gene primers respectively. The recombinant virus was analyzed with Southern blotting and Western blotting. The results indicated that ORF1 and ORF2 gene of PCV2 had been inserted into the genome of TK- /gE- /LacZ+ strain and the expressed ORF1-ORF2 fusion protein could react with PCV2 positive sera. Result of virus titers detection showed the insertion of ORF1 and ORF2 gene did not influence propagation of recombinant virus.
Animals ; Cell Line ; Circovirus ; classification ; genetics ; Gene Transfer Techniques ; Genes, Viral ; Herpesvirus 1, Suid ; genetics ; Open Reading Frames ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Swine

In order to produce relatively large amounts of recombinant human intestinal trefoil factor and assess its biological activity. The expression plasmid pPIC9-hITF containing AOX1 promotor and the sequences of secreting signal peptides was transformed into the yeast cells. Then through selection, positive transformants were cultivated in fermentation basal salts medium in a 5L fermenter to obtain large amount product with low cost. The secreted peptides were then purified by a combination of ionic exchange chromatography and molecular sieve. To verify the product, electrospray mass spectrometry analyses was used to determine the structure of rhITF and Western Blotting was performed to test the immunological activity. Furthermore, the biological activity of the peptide was examined by experiments from cell to tissue. The nucleotide sequence of rhITF was the same as expected. With a 5-L fermenter, 253mg of hITF was isolated at the purity of 96% from 3.5 L of yeast fermentation broth. The expression level for recombinant human ITF in this yeast system was 73.33mg/L. In our study, we provided a way to gain a production among milligram to gram of recombinant human ITF by the use of a yeast expression system. As human ITF are difficult to purify in any significant amount from tissue extraction, the way described may become a valuable tool in obtaining pure peptide for further studies of trefoil peptide function.
Cell Movement ; drug effects ; Epithelial Cells ; cytology ; drug effects ; Fermentation ; Humans ; Intestine, Small ; cytology ; Peptides ; genetics ; metabolism ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Trefoil Factor-2

OBJECTIVETo understand the effects of the IL-1, TNF-alpha cytokines on the pathogenesis of periapical lesions by investigating its gene expression in rat.

METHODSThe model was established by surgically exposing mandibular molar teeth and left open to permit infection from the oral environment. The SD rats were killed at 1, 2, 3, 4 weeks and mandibular molar teeth X-ray were taken. In situ hybridization was used to test IL-1alpha, IL-1beta, TNF-alpha mRNA expression in periapical area and the kinds of positive cells were identified. Using image analysis system analyzed gene expression semi-quantified.

RESULTSIL-1alpha, IL-1beta and TNF-alpha mRNA were all expressed beginning at 1 week, peaked at 3 weeks, and declined somewhat at 4 weeks, but IL-1beta mRNA was expressed at much lower levels with the same kinetics (P<0.01). Most of the staining occurred in areas that had heavy inflammatory infiltrate, fibroblasts, or endothelial cells. There was a statistically significant correlations between the area of periapical lesion and the number of positively stained cells for IL-1alpha and for TNF-alpha (IL-1alpha: r=0.875, P<0.001; TNF-alpha: r=0.858, P<0.001), and between the number of positive cells for IL-1alpha and that of for TNF-alpha (r=0.969, P<0.001).

CONCLUSIONSIL-1alpha and TNF-alpha genes are highly expressed in developing periapical lesions in rat, which supports the hypothesis that these two cytokines play a key role in pulpal and periapical pathogenesis, including the concomitant bone destruction.

Animals ; In Situ Hybridization ; Interleukin-1 ; genetics ; physiology ; Periapical Periodontitis ; etiology ; immunology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; physiology

OBJECTIVETo evaluate the toxicity of photoactivated insecticide K-01 to the larvae of Aedes albopictus and observe the histopathological changes in the larvae.

METHODSThe number of dead larvae was recorded after application of K-01 at different concentrations under different illumination conditions. The content variation of the midgut, malpighian tubules and fat bodies in the larvae was observed microscopically, and the genomic DNA of the larvae was extracted for electrophoresis to identify the target bands.

RESULTSThe maximum larvae-killing effect was achieved with 50 mg/ml K-01 applied under sunlight (100% killing 24 hours after application). Optical microscopic observation of the killed larvae revealed severe damage of the mid-intestinal cells that showed disintegration and elongation. Distinct vacuoles were observed in the fat body cells, in which red droplets were seen to assemble around the cell nuclei. The result of 0.8% agarose gel electrophoresis of the larvae genomic DNA presented typical ladder patterns, suggesting the presence of cell apoptosis.

CONCLUSIONK-01 is an effective photoactivated insecticide.

Aedes ; drug effects ; growth & development ; radiation effects ; Animals ; Insecticides ; toxicity ; Larva ; drug effects ; growth & development ; radiation effects ; Ultraviolet Rays

OBJECTIVETo examine the relationship between effect of vascular endothelial growth factor (VEGF) on epithelial-myofibroblast transition (EMT) of HK-2 cells and changes in expressions of bone morphogenetic protein-7 (BMP-7) and inhibitor of DNA binding/differentiation (Id) 2, Id3.

METHODSThe cultured HK-2 cells were co-treated with transforming growth factor-beta1 (TGF-beta1) (5 ng/ml) and VEGF165 (0.1, 1, 10, 100 ng/ml), or with TGF-beta1 (5 ng/ml) and VEGF receptor-1 neutralized antibody (10 microg/ ml), and were also co-treated with TGF-beta1 (5 ng/ml) and VEGF165 (100 ng/ml) with or without activin receptor-like kinase 6 (Alk6)/Fc Chimera (2 microg/ml, to neutralize endogenous BMP-7) for 48 hours. mRNA and protein expressions of alpha-smooth muscle actin (alpha-SMA), E-cadherin, BMP-7, Id2 and Id3 of HK-2 cells were assessed with double-stain immunocytochemistry, real-time PCR and Western blot respectively.

RESULTSCompared with normal controls, alpha-SMA expression significantly increased, while E-cadherin, BMP-7, Id2, and Id3 mRNA and protein expressions markedly decreased in HK-2 cells treated with TGF-beta1 (5 ng/ml) (P < 0.05). VEGF165 interrupted TGF-beta1 induced alpha-SMA expression in a dose-dependent manner and upregulated BMP-7, Id2 mRNA and protein expressions of the cells (P < 0.05). alpha-SMA expression increased, while E-cadherin, BMP-7, and Id2 expressions decreased further in HK-2 cells co-treated with TGF-beta1 and VEGFR1 antibody compared with normal controls (P < 0.05). When endogenous BMP-7 was neutralized with Alk6/Fc Chimera in the cells co-treated with TGF-beta1 and VEGF165, alpha-SMA expression upregulated (P < 0.05), while Id2 was not changed.

CONCLUSIONSVEGF165 may partially inhibit TGF-beta1-induced EMT of HK-2 cells in vitro. This effect is related to the upregulated expressions of BMP-7 and Id2. Id2 may be upregulated directly by VEGF165, but not related to BMP-7.

Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Cell Differentiation ; Cell Line ; Epithelial Cells ; cytology ; metabolism ; Gene Expression Regulation ; Humans ; Inhibitor of Differentiation Protein 2 ; genetics ; metabolism ; Inhibitor of Differentiation Proteins ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism