【 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.
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 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.
ObjectiveTo observe the changes of macular visual function after myopic foveoschisis (MF) and pars plana vitrectomy (PPV) combined with internal limiting membrane peeling and air filling. MethodsA single-center, retrospective study. From October 2018 to October 2019, 29 MF inpatients (32 eyes) in Tianjin Eye Hospital were included in this study. There were 3 males (4 eyes) and 26 females (28 eyes). The age was 63.00±3.45 years old. Equivalent spherical lens degree was －14.16±2.54 D, and axial length was 29.14±1.04 mm. Among them, 3 patients (3 eyes) had lamellar macular holes. All eyes underwent standard pars plana three-channel 25G PPV combined with internal limiting membrane peeling and air filling. Before surgery and at 1, 3, and 6 months after surgery, macular microperimetry was performed with a macular integrity assessment instrument, and the mean retinal sensitivitie (MS) within 10° of the macula, fovea 2° and 4° fixation rates (P1, P2), 63% and 95% bivariate contour ellipse area (BCEA) were recorded. The comparison of MS, P1, P2, 63%BCEA and 95%BCEA at different times before and after surgery was performed by paired t test; the comparison of fixation stability rate was performed by χ2 test. ResultsCompared with before surgery, there were significant differences in the improvement of MS in affected eyes at 1, 3 and 6 months after surgery (t=－2.208, －3.435,－4.919; P=0.038, 0.002, 0.000). In the pairwise comparison at different times after surgery, only 6 months after surgery and 1 month after surgery were significantly different (P=0.036). Compared with the preoperative P1, P2, 63%BCEA and 95%BCEA, the P1 and P2 of the eyes gradually increased after surgery, while the 63%BCEA and 95%BCEA gradually decreased, however, the difference was not statistically significant (P1: t=－1.595,－1.698,－1.966; P=0.125, 0.104, 0.062. P2: t=－1.622,－1.654,－1.707; P=0.119, 0.112, 0.102. 63%BCEA: t=1.410, 1.409, 1.553; P=0.172, 0.173, 0.135. 95%BCEA: t=1.412, 1.408, 1.564; P=0.172, 0.173, 0.132). Six months after surgery, all the eyes underwent anatomical repositioning of the macular area, and no serious complications such as full-thickness macular hole and macular hole retinal detachment were found. ConclusionsPPV with internal limiting membrane peeling and air filling is an effective and safe method for MF, and the macular function improved significantly within 6 months postoperatively.
Objective To investigate the radiation-sensitizing effects of Ku80 silencing by siRNA interference for A549 lung cancer cells. Methods The sequences of Ku80 siRNA and negative siRNA were chemically synthesized and transfected into A549 lung cancer cells by lipofectamine. RT-PCR and Western bolt analysis were used to determine Ku80 gene expression. The transfected cells in culture dishes were irradiated with X ray at doses of 2 Gy, 4 Gy, 6 Gy, 8 Gy, 10 Gy, respectively. Once all treatments were completed, the cells were processed with the colony formation assay. Results RT-PCR detection showed that Ku80 mRNA levels in A549 lung cancer cells were reduced after transfected with Ku80 siRNA at 24 h, 48 h and 72 h time points. Western blot analysis showed that Ku80 protein content decreased at 48 h and 72 h time points compared with the control group ( P lt; 0. 05 ) . Cloning formation assay indicated that radiosensitivity of A549 lung cancer cells was enhanced after transfected with Ku80 siRNA. Conclusion Ku80 siRNA can effectively inhibit Ku80 gene expression of A549 lung cancer cells, and therefore enhance its radiosensitivity.
Objective To investigate the correlation between OPRM1 A118G gene polymorphism and Eysenck personality type and pain sensitivity. Methods The surgical patients who were transferred from Department of Emergency Medicine to Department of General Surgery of Luzhou People’s Hospital between January 2018 and December 2020 were selected. Before surgery, Eysenck Personality Questionnai (EPQ) was used to investigate the patient’s personality type, and the pain threshold and pain tolerance threshold were determined by electric stimulation instrument. The OPRM1 A118G genotype of peripheral venous blood was detected by polymerase chain reaction-restriction fragment length polymorphism analysis technique. Patients were divided into wild homozygous (A/A) group, mutant heterozygous (A/G) group and mutant homozygous (G/G) group according to the typing results. The general condition, pain sensitivity, EPQ score, difference of Eysenck personality type and correlation between Eysenck personality type and pain sensitivity were analyzed. Results A total of 356 patients were enrolled, including 174 in A/A group, 136 in A/G group and 46 in G/G group. The mutation rate of OPRM1 A118G gene was 32.00%. There were statistically significant differences in pain sensitivity (pain threshold, pain tolerance threshold) and scores of introverted and extraverted, neurotic and dissemble personality types among three groups (P<0.05). There were significant differences in introverted and extraverted and psychotic personality types among the three groups (P<0.05). There were significant differences in pain threshold and pain tolerance threshold among different introverted, extraverted and psychotropic personality types (P<0.05). Conclusion Both OPRM1 A118G gene polymorphism and Eysenck personality type have influence on pain sensitivity, and there is a correlation between them.