Objective: To establish a HepG2 cell line stably overexpressing hsa-mir-122 by transfecting the HepG2 cells with pEZX-hsa-mir-122 vectors,so as to observe the influence of miR-122 on the lipid metabolism in the liver cells. Methods: The constructed pEZX-hsa-mir-122 and pEZX-mir-control plasmids were identified by restriction enzyme digestion and the right clones by sequencing were subjected to sequencing. And the right clones by sequencing were transfected by Lipofectamine™. The transfection efficiency was assessed by green fluorescent protein expression. The stably transfected cell lines were screened by Puromycin. The expression of miR-122 was examined by real-time PCR. The inhibitory function of miR-122 was testified by Western blotting assay. Nile red was used to observe the content of lipid in HepG2 cells stably overexpressing hsa-mir-122. Results: HepG2 cells were successfully transfected with pEZX-hsa-mir-122 and pEZX-mir-control plasmids. The precursor miRNA sequences were integrated into the genome of HepG2 cells stably overexpressing miR-122. Compared with the control HepG2 cells, the HepG2 cells overexpressing miR-122 contained more lipid in the cytoplasm. Conclusion: The miR-122 precursor sequence can intergrate into the genome of HepG2 cells. The increase of miR-122 level in HepG2 cells can result in abnormal metabolism of lipid.

Humans ; Infant ; Male ; Mycobacterium tuberculosis ; Otitis Media ; microbiology ; Tuberculosis

In this study, pEGFP-N1 was chosen as the reporter plasmid and transferred into Ctenopharyngodon idellus kidney (CIK) cells by electroporation, and the optimal electroporation conditions were determined by testing the transfection efficiency with different voltages, pulse times, plasmid amounts, and numbers of shocks. The results showed that the maximum electroporation efficiency was achieved under the following conditions in a 0.2 cm electroporation cuvette containing CIK cells (1.5 x 10(7)/mL, 200 microl): electric voltage 200 V, pulse time 45 ms, plasmid 30 microg, and one electric shock. The total genomic RNA of grass carp reovirus (GCRV) was extracted in this experiment and reversely transcribed into cDNA, which was used to amplify the gene segment of GCRV non-structural protein NS26 using designed specific primers. The PCR product was recombined into pEGFP-N1 vector. The fusion protein EGFP-NS26 was successfully and efficiently expressed in the CIK cells by electroporation, which was confirmed by both fluorescent imaging and Western blot analysis. This experiment laid a foundation for further functional studies of the non-structural protein NS26 of GCRV.
Animals ; Cell Line ; Cyprinidae ; Electroporation ; Fish Diseases ; virology ; Gene Expression ; Kidney ; virology ; Reoviridae ; genetics ; physiology ; Reoviridae Infections ; veterinary ; virology ; Viral Nonstructural Proteins ; genetics ; metabolism

Objective To analyze the distribution and antimicrobial resistance of pathogenic bacteria in urinary tract infection (UTI)so as to provide evidence for appropriate selection of antimicrobial agents in clinical practice. Methods From January 2001 to December 2008 in Shanghai Ruijin Hospital,4683 strains of pathogenic bacteria isolated from urine samples were detected by ATB system;drug susceptibility test was performed with disk diffusion method and pathogenic bacteria distribution and drug resistance was analyzed with WHO NET 5.3 software. Results Among 4683 strains of pathogenic bacteria,most was gramnegative bacilli,accounting for about 77.8%,of which predominant strain was Escherichia coli (68.7%,3217/4683).The predominant strain of gram-positive bacteria was Enterococcus faecalis,accounting for 10.0%(468/4683).Escherichia coli showed hish resistance rotes to ampicillin,piperacillin and compound snlfamethoxazole(SMZ-TMP),which were 76.6%,61.7%and 57.4%respectively,while a low resistance to imipenem,cefoperazone-sulbactam,piperacillin-tazobactam,Enterococcus faecalis showed high resistance rates to erythromycin,gentamicin and levofloxacin,which were 65.8%,43.2%and 31.1%respectively,and were most susceptive to vancomycin and teicoplanin, both with resistance rates of 0. The susceptibility rate of Enterobacteriaceae to imipenem was 100%. From 2006 to 2008, the detection rate of extend-spectrum β-lactamases ESBLs -producing Escherichia coil in outpatient increased year by year, from 28.7% to 43.3% (P＜0.05), whereas no significant change was found in inpatients. The detection rate of (ESBLs)-producing Escherichia coil in inpatients was significantly higher than that in outpatients (P＜0.05).The detection rate of ESBLs-producing Escherichia coil was 23.6%. The antimicrobial resistance rate in elderly patients was significantly higher than that in overall antimicrobial resistance rote (P＜0.05). Conclusions The predominant bacteria of UTI are still gram-negative bacteria, main of which is Escherichia col. Bacteria are resistant to a variety of antibiotics. Approximate selection of antibiotics in clinical practice should be made on the basis of susceptibility test results.

Objective To compare the effect of transthoracic contrast echocardiography (cTTE) with transesophageal contrast echocardiography (cTEE) for detecting right to left shunt (RLS) in patients with patent foramen ovale (PFO). Methods The prospective study was conducted in 29 consecutive patients with PFO who suffered from cryptogenic stroke and/or migraine. Contrast echocardiography was performed in all 29 patients. The cTTE was performed using transducer with second harmonic imaging modality (transmitting frequency 1.7 MHz, receiving frequency 3.4 MHz). The cTEE was performed using transducer with frequency 7 MHz. Ten milliliter saline solution of contrast were rapidly administrated through an antecubital vein. According to whether microbubble (MB) appearing in left atrium after complete opaciifcation of right atrium within the ifrst 3 circles, the results were classiifed by a four-level semi-quantitative categorization:Level 1 (no PFO-RSL), no MB in left atrium; Level 2 (small PFO-RSL) 1-10 MBs; Level 3 (medium PFO-RSL) 10-30 MBs;Level 4 (large PFO-RSL)>30 MBs. Results The total detection rate of PFO-RSL was signiifcant different between cTTE and cTEE (86.2%vs 55.2%,χ2=5.711, P=0.017). In cTTE there were 4 cases at level 1, 1 case at level 2, 5 cases at level 3 and 19 cases at level 4. In cTEE there were 13 cases at leverl 1, 2 cases at level 2, 6 cases at level 3 and 7 cases at level 4. The comparison of semi-quantitative grading derived from cTTE and cTEE was also signiifcant different (Wilcoxon signed ranks test showed Z=-3.789, P=0.000). Conclusions The efifciency in detection of PFO-RLS with cTTE was better than with cTEE. Compared with cTEE, cTTE was easier in practice and brought less discomfort and complications to patients.

Nitrite oxidoreductase(NXR) is the key enzyme responsible for the oxidation of NO2-to NO3-in nitrite-oxidizing bacteria.Since NXR is a dissoluble enzyme,located at the inner side of the membranes of cells,its function is dependent on the electron transfer chain related to membranes.This paper reviews the advances in study on NXR,including the structure,catalysis mechanism and the impact of different factors.New techniques applied in recent studies and research prospects are also presented.

Body fat of 12 male adults were measured by water displacement me-thod(density method) at every morning for 5 successive days. The standard deviation of single observation was 0.29kg calculated by mean residual lung volume method. It was significantly lower than the value (0.5kg) calculated by the ordinary method (p

Objective To identify the clinical characteristics and pathological changes of patients suffered from glomerulonephritis complicating with acute interstitial nephritis (AIN) . Methods Twenty one patients of glomerulonephritis complicating with AIN diagnosed by renal biopsy were retrospectively analyzed . Thirty-five pure AIN patients were selected as controls .Results Glomerulonephritis complicating AIN accounted for 37 .5% of all the AIN cases . Βlactam antibiotics and Chinese herbs were the major causes of AIN . 76 .2% of cases received further examinations due to the elevation of serum creatinine (Scr) during their follow-up of kidney injuries or during routine exams for all kinds of discomforts . Pathological features of AIN were also detected besides glomerular leisions . The impairments of renal interstitia were severe than those of the glomeruli . Eosinophil in the renal interstitia was an important indicator for the diagnosis of AIN .The renal function returned to normal or baseline in 64 .7% of the patients of glomerulonephritis complicating with AIN whose follow-up data were available . The median period for renal function restoration was 150 days (compared with 60 days in pure AIN) . But there were no significant differences between these two groups as for the rate of irreversible renal insufficiency during a follow-up period of 2 years . Conclusions Symptoms of AIN in patients of glomerulonephritis complicating with AIN tend to be masked by their glomerular diseases . Renal biopsy is of most importance for the diagnosis . Early diagnosis and treatment leads to satisfactory prognosis .

A total of 152 strains of endophytic fungi were isolated from the barks of Eucommia ulmoides in three regions (Lueyang country, Zunyi country, Cili country). Based on morphological characteristics and analysis of ITS sequences, these strains were identified into 8 genera. Thereinto Phomopsis, Diaporthe and Alternaria were common genera to Eucommia barks from different sites. But the dominant genus was different: Alternaria was the dominant genus in the barks from Cili country, and Phomopsis was the dominant genus from Zunyi country, then Diaporthe was the one from Lueyang country. According to the similarity coefficient, the composition of the endophytic fungi was distinctly different between the barks from three sites. The diversity and species richness in Lueyang country and Cili country were found higher than those in Zunyi country. The evenness of endophytic fungi was 0.936 5 in Lueyang county, which was higher than 0.737 1 or 0.641 0 in Cili county or Zunyi county, respectively. After phylogenic analysis and calculating the genetic distances of typical strains belong to Phomopsis and its perfect stage--Diaporthe, there was very high genetic diversity in the two genera from our study. In conclusion, the community structure and diversity of endophytic fungi were significant different in Eucommia barks from the three habitats.
DNA, Fungal ; genetics ; DNA, Intergenic ; genetics ; Ecosystem ; Endophytes ; classification ; physiology ; Eucommiaceae ; microbiology ; Fungi ; classification ; genetics ; physiology ; Phylogeny ; Plant Bark ; microbiology

AIM: To explore the long-term efficacy of Q-factor guided laser epithelial keratomleusis ( LASEK ) for myopia and astigmatism with positive Q-factor.
METHODS: There were 158 eyes which were myopia and astigmatism with positive Q- factor taken in two groups randomly: 86 eyes accepted Q - factor guided LASEK as observation group and 72 eyes accepted routine LASEK as control group. The difference between the two groups about all data was similar. The uncorrected visual acuity ( UCVA ) and the best corrected visual acuity ( BCVA ) as well as diopter, ocular tension, corneal topography, Keratometry value K, aspherical factor Q, Higher-order aberrations ( HOA ) , corneal thickness by ultrasound and, contrast sensitivity ( CS ) , Haze were examined and compared before and after surgery. All the cased were followed up for 14d, 1, 3, 6, 12mo. And there were no statistical difference among the data before surgery.
RESULTS: After 12mo there were no statistical difference between the two groups about UCVA and BCVA. But the safety index of observation group was 1.10, that of control group was 1. 07. The validity index of observation group was 1. 06, that of control group was 0.99. The HOA of observation group was 0. 45±0. 17μm, and that of control group was 0. 72±0.25μm, there was statistically significant difference (t=-8. 193,P=0. 000). Q factor of observation group was 0. 41±0. 17, that of control group was 0. 77±0. 22, there was significant difference (t=11. 377,P = 0. 028). The contrast sensitivity of 3mo post surgery of patients in the observation group returned to the level of before surgery. But in the control group the contrast sensitivity of the patients did not returned until 6mo.
CONCLUSION:Q-factor guided LASEK for myopia and astigmatism with positive Q-factor is stable, safe and effective. The operation allow for reducing the high order aberrations, maintaining the most asphericity of cornea, saving more in corneal tissue, which cause faster recovery of contrast sensitivity, less haze and better visual quality.