【 Abstract 】 Objective To observe the effect of disruption of hypoxia inducible factor-1 α (HIF-1 α ) pathway by small hairpin RNA (shRNA) on chemosensitivity of human hepatocellular carcinoma (HCC) cells and to reveal the correlative mechanisms. Methods Plasmid of pshRNA-HIF-1α was transfected into HepG2 cells by lipofectamine. HepG2/pshRNA-HIF-1α (HepG2/pshRNA) cell lines were obtained by selection of HepG2 cells in G418. Meanwhile, plasmid of empty vector (pHK) was transfected as a control (HepG2/pHK). The mRNA and protein expression levels of HIF-1α and mdr1 were investigated by RT-PCR and Western blot respectively. Using CoCl2 to simulate the hypoxia condition, growth inhibition and apoptosis rates of HepG2 cells under different dosages of chemotherapeutic agents (adriamycin) were measured by MTT assay and flow cytometry (FCM) ． ResultsCompared with HepG2/pHK cells, the mRNA and protein expression levels of HIF-1αand mdr1 were obviously down-regulated in HepG2/pshRNA cells. At the same time, the proliferation inhibition and apoptosis rates were evidently increased after transfection with pshRNA-HIF-1α(P＜0.05)，which decreased the expression of HIF-1αto 82.18% at mRNA level and 75.51% at protein level. There was no significant effect of transfection pHK (Pgt;0.05). Conclusion These data demonstrates that HIF-1α interference by shRNA increased the sensitivity of HCC chemotherapy and the reversal of multidrug resistance, which may be done by down-regulating the transcription of mdr1 and the translation of P-gp. Blocking HIF-1αin HCC cells may offer an new avenue for gene therapy．
摘要:目的: 金黄色葡萄球菌(金葡菌)的感染近年来已成为医院内的主要致病菌,而其耐药性也呈逐渐升高的趋势,为了解该菌在我院的感染和耐药情况,为临床合理使用抗生素提供科学依据。 方法 : 用经典生理生化鉴定方法,对各种临床标本主要来源于痰液和各种伤口脓液标本分离到的102株金葡菌进行生物学特性及药敏试验。 结果 : 从我们医院2007年5月至2009年8月所分离出来的102株金葡菌中青霉素耐药性8923%,氨苄青霉素耐药率为9385%,没有发现万古霉素耐药菌。 结论 : 除万古霉素外,耐药率较低的依次是利福平、苯唑青霉素、环丙沙星、呋喃妥因、阿米卡星、磺胺甲基异恶唑、红霉素,而青霉素G、氨苄青霉素、四环素耐药性情况非常严重,并且多重耐药,耐药性强,应引起临床的高度重视。Abstract: Objective: To analyze the bionomics and antimicrobial susceptibility of staphylococcus aureus, which was the main pathogenic bacterium with high drug tolerance in our hospital, in order to provide the rational use of antibiotics. Methods : Samples of one hundred and two staphylococcus aureus cases from sputamentum and pus were evaluated by classic physiology and biochemistry methods to test the bionomics and antimicrobial susceptibility. Results : The drug resistance rate to penicillin, penbritin and vancomycin was 8923%, 9385% and 0, separately. Conclusion : Besides vancomycin, the drug resistance rate of rifampicin, oxazocilline, ciprofloxacin, furadantin, amikacin, sulfamethoxazole and sulfamethoxazole increased one by one. The resistance to penicillin G, penbritin and tetracycline was serious, including multidrug resistant, which should be paid highly attention.
We have studied the radiosenstivity of retinoblastoma cell [inc: HXO Rb~4,and found that the ceil growth reduced markedly after being treated by 3GyT-ray. From both clone for mation method and MTT assay,we identify that HXO-Rb44 cell is radiosensitive to T-ray.Oxygen can increase the radiosensitivity of HXO-Rb44 cell, but decrease the repair of sublethal damage.Oxygen enchaneement ratio(OER)is 2.77~3.01. （Chin J Ocul Fundus Dis,1994,10:217-219）
Objective To determine the correlation between central macular thickness (CMT) and the visual function in patients iwht macular edema (ME). Methods The clinical data of 42 eyes of 40 patients with ME which were examined by optical coherence tomography (OCT) and microperimetry (MP-1) were retrospectively analyzed. In 40 patients (42 eyes), diabetic ME (DME) was in 27 eyes，branch retinal vein occlusion was in 11eyes, and central retinal vein occlusion was in 4 eyes. All of the eyes had undergone OCT，MP-1 and best-corrected visual acuity (BCVA) test. Central macular thickness (CMT) was measured by fast macular scans using OCT. Retinal sensitivity (MS) and fixation patterns were evaluated by Mp-1. The position was chosen :2 disc diameters (DD) temporal to the disc and one third of a DD inferior to the centre of the disc. Results The correlation between CMT and BCVA is not significant (r＝－0.429，P=0.069) as well as the correlation between CMT and MS (r＝－0.433，P=0.058). The difference of CMT between the unstable and stable group was significant (F=3.262, P=0.039）. The difference of CMT between the central fixation group and preferred retinal locus (PRL) group was significant (F=3.173, P=0.044). Conclusions BCVA and MS have no significant correlation with CMT. When CMT increases, the fixation stability decreases, fixation location.changes, and PRL occurs.
Objective To investigate the effects of X-ray dose on the expressions of microRNA-221 （miR-221） and phosphatase and a tensin homolog deleted from chromosome10 （PTEN） in human colorectal carcinoma （CRC） cells. Methods Human CRC-derived cell line， Caco2， was cultured conventionally. The cells were divided into five groups and exposed to different doses of X-ray （0， 2， 4， 6， and 8 Gy） respectively. The total RNA and protein of the Caco2 cells were extracted after irradiation， and the miR-221 and PTEN mRNA expressions were detected by real-time RT-PCR.Moreover， the protein alteration of PTEN in Caco2 cells was detected by Western-blot analysis. Results The radiation dose of X-ray significantly affected the expressions of miR-221 and PTEN protein in human Caco2 cells in a dose-depen-dent manner. Moreover， the miR-221 expression level was up-regulated gradually with the increase of irradiation dose， on the contrary， the PTEN protein expression level was down-regulated gradually （P＜0.01）. Conclusion The radiation dose can affect the miR-221 and PTEN protein expression pattern in CRC cells.
Objective To explore the microbiological etiology and antibiotic susceptibility of periopertive urinary tract infection (UTI) in patients undergoing hip or knee arthroplasty, so as to provide recommendations for antibiotic treatment. Methods A retrospective review was conducted for patients with perioperative UTI who underwent hip or knee arthroplasty between January 1st, 2013 and October 1st, 2015. Microbiological data and antibiotic susceptibility of bacteria were analyzed. Results A total of 117 strains of bacteria were identified, including 11 types of species. Among the organisms cultured, 86.3% (101 strains) were gram-negative bacteria, in which Escherichia coli was the most common causative organism (70.9%, 83 strains), followed by Klebsiella species (7.7%, 9 strains) and Proteus mirabilis (3.4%, 4 strains). And among the gram-positive bacteria detected, the proportion of Enterococcus faecalis and Feces Enterococcus was 6.8% (8 strains) and 3.4% (4 strains), respectively. The bacteria showed highly resistance to cephalosporins, quinolones and sulfonamides, but showed high sensitive to nitrofurantoin, carbopenems, the enzyme inhibitor complex and aminoglycoside antibiotics. Conclusions There is a diversity of bacteria involved in UTI, and the top 3 pathogens are Escherichia coli, Enterococcus faecalis and Klebsiella species. The resistance rate is high, and nitrofurantoin, amilacin, piperacillin-tazobactam, cefoperazone-sulbactam are the recommended antibiotics to treat the UTI, but the antibiotic should be adjusted according to susceptibility results.
Although anaphylaxis induced by vitamin K1 seldom happens, 4 allergic cases were observed in the patients we treated recently who were given intramuscular injection of vitamin K1 before renal biopsy. To provide the best clinical evidence, we searched MEDLINE （-May 2005） and evaluated the studies. The studies were only case reports and retrospective reviews which showed the anaphylaxis were mainly allergic dermatitis with different manifestation and reaction time. The serious reactions such as allergic shock was very rare. We conclude that although vitamin K1 anaphylaxis is rare, strict indications should be followed and the drug surveillance on adverse events should be strengthened.
ObjectiveTo strengthen the understanding to hypersensitivity pneumonitis and make early diagnosis and standard treatment by analyzing the clinical features, the diagnosis and treatment of a patient diagnosed as hypersensitivity pneumonitis presenting as progressive fibrotic interstitial lung disease in combination with literature review.MethodsThe diagnosis and treatment process and relevant clinical data of the patient were analyzed retrospectively and literatures were reviewed. Based on 282 relevant literatures, the diagnostic methods, treatment and prognostic factors of hypersensitivity pneumonitis were summarized.ResultsThe patient, female, 45 years old, self-employed, was admitted to the hospital due to "cough, sputum for 9 years, dyspnea for more than 6 months" without obvious extrapulmonary clinical manifestations. Creaks at the bottom of the lungs and clubbed toes were found through physical examination. High-resolution computed tomography indicated that the main manifestations were ground glass and grid-like shadows, presenting characteristics of interstitial pneumonia. The pulmonary lesions aggravated gradually, and part of the lung lobe presented honeycomb lung at the time of diagnosis. Eight years ago, she performed fibrobronchoscopy in other hospital revealing an increased lymphocyte proportion (39%) in bronchoalveolar lavage fluid. Lung function suggested very severe restrictive ventilation dysfunction. She was given prednisone for a short time, and the drug was stopped by herself with uncertain efficacy. Later, she performed frozen lung biopsy suggesting peribronchiolar metaplasia interstitial broadening with lymphocytic infiltration, without granuloma or fibroblast lesions. No obvious abnormity was seen in makers of infections and immunology. At this point, the patient was clearly diagnosed as hypersensitivity pneumonitis. Meanwhile, the patient's pulmonary lesions were still mainly made of ground glass and plaques, partly with changes like honeycomb. Poor response was obtained with anti-inflammatory treatment of prednisone for 3 months and anti-fibrosis treatment of pirfenidon for more than 2 months. Literature review resulted in 87 patients, including 39 males (44.82%) and 48 females (55.18%), with an average age of (47.0±18.4) years. Seventy patients (80.46%) had clear allergens. 3.45% patients' lymphocyte percentage in bronchoalveolar lavage fluid ranged from 20% to 39%, and 19.54% patients' lymphocyte percentage was more than 40%. 4.60% of the patients achieved remission through detachment from allergens; 71.27% of the patients achieved remission with glucocorticoid treatment, 14.94% improved with glucocorticoid treatment, and 1.15% died; 6.89% of the patients achieved remission after receiving anti-allergy therapy. Patients with chronic hypersensitivity pneumonia were found with poor prognosis.ConclusionsIn patients with diffuse interstitial pulmonary fibrosis with unknown cause, transbronchoscope freezing lung biopsy as early as possible is a feasible method for early diagnosis and improving prognosis. Patients with hypersensitivity pneumonitis with a long course of pulmonary fibrosis have a poor response to glucocorticoid and other classic treatments, and most of them have a poor prognosis.
ObjectiveTo evaluate the predictive value of the high-sensitivity cardiac troponin I (hs-cTnI) in patients with acute pulmonary embolism (APE). MethodsIn a retrospective cohort study,272 consecutive patients with APE were reviewed and the 30-days death and in-hospital adverse events were evaluated. The patients were classified according to hs-cTnI value into a high hs-cTnI group and a low hs-cTnI group. The simple pulmonary embolism severity index (sPESI) was used for clinical risk determination. The adverse event was defined as intravenous thrombolytic therapy,noninvasive ventilator support to maintain oxygen saturation >90% and suffered with severe complications. The correlations of hs-cTnI with sPESI score,30-days adverse events and mortality were analyzed. The Kaplan-Meier curves and the log-rank test were used to compare time-to-event survival. Stepwise multivariate logistic regression analysis models were used to determine the incremental prognostic value of sPESI score and hs-cTnI. ResultsThe incidence of 30-day death (6.1%),renal failure (14.6%),bleeding (13.4%) and thrombolytic therapy (7.9%) were higher in the high hs-cTnI group than those in the low hs-cTnI group (P values were 0.009,<0.001,0.018 and 0.003,respectively). The patients with sPESI ≥1 and low hs-cTnI had greater free adverse events survival (P=0.005). hs-cTnI provided incremental predictive value for in-hospital adverse events,beyond the sPESI score (P<0.001). Conclusionhs-cTnI has excellent negative predictive value of APE prognosis,especially when used combined with sPESI score.