Objective To study the apoptosis of pancreatic cancer cells induced by a retroviral vector containing antisense human telomerase RNA gene. Methods Sense and antisense telomerase RNA genes were transfected into Can-pan-2 cell line by electroporation,and selected with G418 to get stable transfection and retrovirus yielding cell lines. We transfected Can-pan-2 cell with concentrated retroviral supernatant,and selected with G418 to get stable transfection cell lines,The expression of the aim gene in Can-pan-2 cell was confirmed through PCR,while TRAP-PCR-ELISA was used to determine the telomerase activity,and immunofluorescence was adopted to determine reproductive activity of the infected cells,flow cytometry(FCM)was used to detect cell apoptosis.ResultsThe titer of sense and antisense recombinant retrovirus reached 0.95?106 CFU/mL and 1.1?106 CFU/mL respectively,PCR demonstrated that the telomerase RNA genes were successfully intergrated into the target cells’ genome,The cell growth and telomerase activity of Can-pan-2 were significantly inhibited and apoptosis occurred after antisense virus infection. Conclusion Antisense human telomerase RNA may inhibite telomerase activity of pancreatic cancer cells and promote apoptosis of the cells.

BACKGROUND: β-amyloid has been proved to be capable of inducing cell apoptosis and play a vital role in Alzheimer disease (AD).OBJECTIVE: To probe into olanzapine's protective effect and mechanism of PC12 cell apoptosis induced by β-amyloid 25-35.SETTING: Department of Neurology, Southern Building of the General Hospital of Chinese PLA.DESIGN: Randomized design.MATERIALS: This experiment was carried out in the Neuropsychopathic Research Institute, Medical College of the University of Saskatchewan(Canada), between May 2002 and March 2003.METHODS: P12 cells were cultured with RPMI1640 culture medium.100 μL cell suspension was inoculated in each well of 96-well culture plate, and 5 mL suspension was inoculated in 25 cm2 culture bottle covered with collagen and cultured for 24 hours, then with additional 50 μmol/L and 100 μmol/L olanzapine, respectively, for 24 hours, and β-amyloid 25-35 of different concentrations (0.01 μmol/L, 2 μmol/L and 20 μmol/L) for 24 hours. PC12 cell apoptosis was induced by β-amyloid 25-35 in 96-well culture plate and cells were harvested to assay their survival rate with MTF colorimetric assay. PC12 cells in 25-cm2 culture bottles were also harvested to detect the effect of olanzapine on Bax and caspase-3 expression in PC12 cells using Western blot assay.MAIN OUTCOME MEASURES: ① Cell survival rate; ② the expression of Bax and caspase-3 in PC12 cells.RESULTS: ① Cell survival rate: cell activity was found declined from 75% to 35% in PC12 cells induced by β-amyloid 25-35, but obviously increased in PC12 cells due to pretreatment with olanzapine of 50 μmol/L and 100 μmol/L. ② Olanzapine's effects on Bax expression in PC12 cell apoptosis induced by β-amyloid 25-35: Bax expression increased in PC12cells due to exposure to β-amyloid 25-35 of 0.01 μmol/L, 2 μmol/L and 20 μmol/L, but it could be suppressed if pretreated with olanzapine of 50 μmol/L. ③ Effect of olanzapine on caspase-3 expression in PC12apoptotic cells induced by β-amyloid 25-35: There was no change in PC12cells induced by 0.001 μmol/L or 0.01 μmol/L of β-amyloid, as well as in PC12 cells pretreated with 50 μmol/L olanzapine. However, caspase-3 expression obviously increased in PC12 cells exposed to 2 μmol/L and 20 μmol/L of β-amyloid 25-35, and it could be suppressed by pretreatment with 50 μmol/L of olanzapine.CONCLUSION: ① β-amyloid 25-35 can induce the high expression of cell apoptosis related Bax and caspase-3 in vitro cultured PC12 cells. ②Olanzapine can reduce the expression, thus enhancing the survival rate of PC 12 cells.

BACKGROUND: Many studies have indicated that amyloid beta-protein (Aβ) plays an important role in the pathophysiology of Alzheimer disease (AD), the reduction of production of Aβ can slow down the deterioration of AD, so to reduce Aβ production could become an important therapeutic target in AD. Many AD patients present behavioral disturbance and psychotic symptoms, and are treated with antipsychotics. Olanzapine and quetiapine can significantly improve the clinical global impressions(CGI) severity-of-Alzheimer scores, clinical studies suggest that early and prolonged intervention can improve long-term outcome.OBJECTIVE: To investigate the effect of olanzapine and quetiapine on the secretion of Aβ42 in Swedish amyloid precursor protein(APP) gene and presenilin 1 gene transfected murine N2a neuroblastoma cells.DESIGN: A completely randomized controlled trial based on N2a cells.MATERIALS: Setting was at Neuropsychiatry Research Institute of Medical College, University of Saskatchewan. The murine N2a and double transfected N2a cell was provided by department of neurology and neuroscience, Cornell university medical college.INTERVENTIONS: The double transfected murine N2a neuroblastoma cells were treated for 24 hours with 200 μmol/L olanzapine and 50 μ mol/L quetiapine respectively, then intracellular and extrocellular levels of Aβ were determined. The MTT assay was used to determine cell viability; the BCA assay was used to determine the protein content of cells; the western blot analysis was used to determine the APP expression; and the Enzyme-Linked-Immuno-Sorbent Assay(ELISA) was used to determine the Aβ produced by double transfected murine N2a neuroblastoma cells.MAIN OUTCOME MEASURES: The levels of intracellular and extracellullar Aβ 42 secreted by double transfected murine N2a neuroblastoma cells were detected using ELISA.RESULTS: The double transfected N2a cells produced more APPs than the naive N2a cells. The extracellular Aβ[ (4.78 ± 0.54) nmol/L] of cells treated with olanzapine decreased significantly comparing to the vehicle [(7.69±0.62) nmol/L] (t=3.52, P ＜0.05); and theextracellular Aβ[ (4. 09 ±0. 18) nmol/L] of cells treated with quetiapine decreased significantly comparing to the vehicle[ (7.50 ±0.50) nmol/L] ( t =5.61,P ＜ 0.05) . The intracellular Aβ of cells treated with olanzapine did not change significantly conpared with the vehicle ( P ＞ 0.05 ); the intracellular Aβ of cells treated with quetiapine did not change significantly compared with the vehicle ( P ＞ 0.05 ).CONCLUSION: The result suggests that olanzapine and quetiapine can decrease the production of Aβ42 in double transfected murine N2a neuroblastoma cells and clinically may be helpful in slowing down the progression of AD by decreasing the extrocellular secretion of Aβ42.

Objective:To observe the etiological distribution, clinical presentations and imaging features of pulmonary mycosis that is diagnosed by pathology.Methods:The etiological distribution, clinical presentations and imaging features of patients with pulmonary mycosis, who were collected in the Affiliated Hospital of Jining Medical University from January 2018 to July 2020, were retrospectively analyzed. The diagnosis of all the patients were confirmed by pathological examination, of lung or bronchi tissue that were obtained through operation, bronchoscope or percutaneous lung puncture biopsy.Results:There were 26 patients' (60.47%, 26/43) pathological specimens were obtained by operation, 14 cases (32.56%, 14/43) were obtained by bronchoscope, and 3 cases (6.98%, 3/43) were obtained by percutaneous lung puncture biopsy. Of the 43 patients who were diagnosed pulmonary mycosis by pathology, 27 patients (62.79%, 27/43) suffered from pulmonary aspergillosis, 11 patients (25.58%, 11/43) suffered from pulmonary cryptococcosis, 3 patients (6.98%, 3/43) suffered from pulmonary mucormycosis, and 2 patients (4.65%, 2/43) suffered from pulmonary candidiasis. There were 27 patients (62.79%, 27/43) with pulmonary fungal disease complicating risk factors of fungal infection, including diabetes mellitus (23.26%,10/43), malignant tumor (16.28%, 7/43), bronchiectasis (9.30%, 4/43), hepatitis B virus (HBV) carrier (6.98%, 3/43), taking glucocorticoids (4.65%, 2/43), pulmonary tuberculosis (4.65%, 2/43), and chemotherapy following colon carcinoma operation (2.33%, 1/43). The common clinical presentations included cough (55.81%, 24/43), expectoration (48.84%, 21/43), hemoptysis (37.21%, 16/43), fever (20.93%, 9/43), gasping (18.60%, 8/43), chest pain (16.28%, 7/43), and hoarseness (3.13%, 1/43). Imaging features of chest included lung nodes in 20 cases (46.51%, 20/43), vascular welt sign in 12 cases (27.91%, 12/43), exudative process in 10 cases (23.26%, 10/43), lung mass or consolidation in 8 cases (18.60%, 8/43), cavitary lesions in 7 cases (16.28%, 7/43), thicken bronchus wall and narrow lumina in 6 cases (13.95%, 6/43), air crescent in 5 cases (11.63%, 5/43).Conclusions:The pulmonary aspergillosis and cryptococcosis are mainly in pulmonary mycosis diagnosed by pathology. The common complications are diabetes mellitus and malignant tumor. The common clinical presentations are cough, expectoration, and hemoptysis. The main imaging features of chest are lung nodes and vascular welt sign can be found in most of pulmonary cryptococcosis.

Objective To make a cross-sectional assessment of the morbidity of lower extremity arterial disease (LEAD) in inpatients with type 2 diabetes mellitus (T2DM) and to analyze its risk factors,thus providing evidence for its clinical prevention.Methods We enrolled 664 inpatients with T2DM from June 2012 to June 2013 and collected clinical data,including age,gender,duration of diabetes,body mass index,smoking,fasting & postprandial blood glucose levels,glycosylated hemoglobin,serum lipids,renal function,fibrinogen,neck ultrasonography,lower extremity vascular ultrasound,ankle brachial index and treatment records.Logistic multiple regression analysis was conducted to identify risk factors for LEAD.Results A total of 247 cases met the diagnostic criteria for LEAD,with morbidity reaching to 37.2％.The percentages of morbidity in patients with different durations of diabetes were:23.12％ (≤ 5 years),27.95％ [(5 10) years],38.71％ [(1015) years],51.16％ [(15-20) years],62.34％ (≥ 20 years).The differences were statistically significant (P＜0.05).Of the patients in the LEAD group,73.2 ％ were treated with antihypertensive medications and 54.6％ were treated with statins.The goal attainment rates for total cholesterol,lowdensity lipoprotein cholesterol,high-density lipoprotein cholesterol and triglycerides were 56.3％,39.3％,47.4％ and 61.5％,respectively,in the LEAD group and 45.1％,34.5％,35％,and 49.4％,respectively,in the non-LEAD group.With the exception of the rates for low density lipoprotein cholesterol,the rates between the two groups are statistically significant (P＜0.05).Significant differences in age,BMI,blood pressure,coronary heart disease,cerebrovascular disease,carotid intima-media thickness,carotid artery plaque,and carotid artery stenosis were also observed between the two groups (P＜0.05 for all parameters).Logistic multiple regression analysis revealed that age,history of diabetes,cerebrovascular disease,carotid artery plaque,and carotid artery stenosis were risk factors for LEAD.Conclusions The morbidity of LEAD is 37.2％ in type 2 diabetic patients.Age,history of diabetes,cerebrovascular disease,carotid artery plaque,and carotid artery stenosis are risk factors for LEAD,while traditional risk factors for atherosclerosis,including hypertension,levels of cholesterol and low-density lipoprotein cholesterol,smoking,and non-drug intervention,are risk factors for LEAD in type 2 diabetic patients.

OBJECTIVE To investigate the condition of drug-resistant genes in MRSA and MSSA. METHODS The drug-resistant genes mecA,ermA/B/C,aac(6′)/aph(2″),aph(3′)-Ⅲ,ant(4′,4″) and tetM of MRSA and MSSA were detected by polymerase chain reaction(PCR). RESULTS The 5 kinds of drug-resistant genes,such as mecA,ermA/B/C,aac(6′)/aph(2″),(aph(3′)-Ⅲ) and tetM were positive in MRSA. CONCLUSIONS MRSA is a multi-resistant pathogen.

A simple and rapid liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was de-veloped and validated for simultaneous determination of acetaminophen and oxycodone in human plasma. Acetaminophen-d4 and oxycodone-d3 were used as internal standards. The challenge en-countered in the method development that the high plasma concentration level of acetaminophen made the MS response saturated while the desired lower limit of quantification (LLOQ) for oxycodone was hard to reach was well solved. The analytes were extracted by protein precipitation using acetonitrile. The matrix effect of the analytes was avoided by chromatographic separation using a hydrophilic C18 column coupled with gradient elution. Multiple reaction monitoring in positive ion mode was performed on tandem mass spectrometer employing electrospray ion source. The calibration curves were linear over the concentration ranges of 40.0–8000 ng/mL and 0.200–40.0 ng/mL for acetaminophen and oxycodone, respectively. This method, which could contribute to high throughput analysis and better clinical drug monitoring, was successfully applied to a pharmacokinetic study in healthy Chinese volunteers.

Objective:To evaluate the effect of paclitaxel on the mast cell-CCL2-macrophage axis in rats with pulmonary hypertension.Methods:Thirty SPF-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 180-220 g, were divided into 3 groups ( n=10 each) using a random number table method: control group (group C), pulmonary hypertension group (group PH), and paclitaxel group (group PTX). The model of pulmonary hypertension was established by subcutaneous injection of monocrotaline 60 mg/kg in rats.At 25 days after establishing the models, paclitaxel 2 mg/kg was injected via the tail vein once every four days, for 4 times in total in group PTX.The equal volume of normal saline was injected in the remaining 2 groups.The mean pulmonary artery pressure (mPAP) was performed at 40 days after establishing the model.The heart was removed and dried, the right ventricle (RV) and left ventricle plus ventricular septum (LV+ S) was weighed, and the Fulton index [RV/(LV+ S)] was calculated.The inferior lobe of left lung was taken, the ratio of media wall thickness of pulmonary vessels was calculated by HE staining, the number of Tryptase + , CD68 + , CD163 + , and Ki67 + cells was recorded by immunohistochemistry, the mean value was calculated, the percentage of Ki67-positive cells in blood vessels was recorded, and the proportion of muscularized blood vessels was calculated.The content of CCL2 was measured by enzyme-linked immunosorbent assay, and the expression of cleaved caspase-3 and Cyclin D1 was detected by Western blot. Results:Compared with group C, the mPAP, Fulton index, ratio of media wall thickness, proportion of muscularized blood vessels, the number of Tryptase + , CD68 + and CD163 + cells and percentage of Ki67 + cells were significantly increased, and the expression of cleaved caspase-3 was down-regulated in PH and PTX groups ( P<0.05), the expression of Cyclin D1 was significantly up-regulated in group PH ( P<0.05), and no significant change was found in group PTX ( P>0.05). Compared with group PH, the mPAP, Fulton index, ratio of media wall thickness, percentage of muscularized blood vessels, the number of Tryptase + , CD68 + and CD163 + cells and percentage of Ki67 + cells were significantly decreased, the expression of CCL2 and Cyclin D1 was down-regulated, and the expression of cleaved caspase-3 was up-regulated in group PTX ( P<0.05). Conclusion:The mechanism by which paclitaxel alleviates pulmonary hypertension is related to inhibiting the mast cell-CCL2-macrophage axis in rats.

Objective To evaluate the efficacy of chemoradiotherapy alone and prognostic factors for locally advanced rectal cancer. Methods The clinical data of 47 patients with locally advanced rectal cancer who were admitted to our hospital and mostly treated with chemoradiotherapy alone from 2003 to 2010 were retrospectively analyzed. Three of the patients received radiotherapy alone. The Kaplan?Meier method was used to estimate overall survival (OS), progression?free survival (PFS), and distant metastasis?free survival ( DMFS ) rates, and the log?rank test was used for survival difference analysis and univariate prognostic analysis. The Cox regression model was used for multivariate prognostic analysis. Results In all patients, the 3?and 5?year OS rates were 53?2% and 33?2%, respectively, while the 3?and 5?year PFS rates were 37% and 31%, respectively. During the follow?up, 15 patients (32%) had local progression with PFS of 1?60 months (median PFS, 14 months);23 patients (49%) had distant metastasis with DMFS of 2?60 months ( median DMFS, 17 months) . Patients treated with high?dose radiotherapy had significantly lower 3?and 5?year local progression rates than patients treated with medium?dose radiotherapy ( 11% vs. 54%;11%vs. 57%;P=0?004). After chemoradiotherapy, 9 patients (19%) had clinical complete response (cCR), and the 3?and 5?year OS and PFS rates in those patients were all 8/9. The univariate analysis indicated that tumor distance from the anus and cCR were influencing factors for prognosis ( P= 0?026;P= 0?000 ) . However, the multivariate analysis showed that cCR was the only influencing factor for survival ( HR=12?24;95% CI, 1?64 ?91?29;P= 0?015 ) . Conclusions Chemoradiotherpay or radiotherapy alone is effective and safe in the treatment of patients with locally advanced rectal cancer who have to give up surgery or have unresectable tumors. High?dose radiotherapy may improve local control rate. Complete response to chemoradiotherapy predicts satisfactory treatment outcomes.

Objective To explore the risk factors for lower extremity amputation in patients with diabetic foot.Methods The clinical data of 1 771 patients with diabetic foot at the Air Force General Hospital of PLA from November 2001 to April 2015 were retrospectively analyzed.The patients were divided into the non-amputation and amputation groups.Within the amputation group , subjects were further divided into the minor and major amputation subgroups.Binary logistic regression analyses were used to assess the association between risk factors and lower extremity amputation.Results Among 1 771 patients with diabetic foot , 323 of them ( 18.24%) were in the amputation group ( major amputation: 41; minor amputation:282 ) and 1 448 ( 81.76%) in the non-amputation group.Compared with non-amputation patients, those in the amputation group had a longer hospital stay and higher estimated glomerular filtration rate(eGFR)levels.Fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c), C-reaction protein (CRP), ESR, ferritin, fibrinogen and WBC levels of the amputation group were higher , while hemoglobin albumin, transferrin, TC, TG, HDL-C and LDL-C were lower than those of the non-amputation group (all P<0.05 ).The proportion of hypertension ( 52.48% vs 59.98%) , peripheral vascular disease ( PAD ) (68.11% vs 25.04%), and coronary heart disease (21.33% vs 28.71%) were different between the amputation and non-amputation groups (all P<0.05).Multivariable logistic regression analyses showed that Wagner′s grade , PAD and CRP were the independent risk factors associated with lower extremity amputation in hospitalized patients with diabetic foot.Conclusion Wagner′s grade, ischemia of lower limbs and infection are closely associated with amputation of diabetic foot patients.