Hospitalization costs of injury in elderly population in China: a quantile regression analysis.

BACKGROUND: Trauma in the elderly is gradually growing more prevalent as the aging population increases over time. The purpose of this study is to assess hospitalization costs of the elderly trauma population and analyze the association between those costs and the features of the elderly trauma population. METHODS: In a retrospective analysis, data on trauma patients over 65 who were admitted to the hospital for the first time due to trauma between January 2017 and March 2022 was collected from a tertiary comprehensive hospital in Baotou. We calculated and analyzed the hospitalization cost components. According to various therapeutic approaches, trauma patients were divided into two subgroups: non-surgical patients (1320 cases) and surgical patients (387 cases). Quantile regression was used to evaluate the relationship between trauma patients and hospitalization costs. RESULTS: This study comprised 1707 trauma patients in total. Mean total hospitalization costs per patient were ¥20,741. Patients with transportation accidents incurred the highest expenditures among those with external causes of trauma, with a mean hospitalization cost of ¥24,918, followed by patients with falls at ¥19,809 on average. Hospitalization costs were dominated by medicine costs (¥7,182 per capita). According to the quantile regression results, all trauma patients' hospitalization costs were considerably increased by length of stay, surgery, the injury severity score (16-24), multimorbidity, thorax injury, and blood transfusion. For non-surgical patients, length of stay, multimorbidity, and the injury severity score (16-24) were all substantially linked to higher hospitalization costs. For surgical patients, length of stay, injury severity score (16-24), and hip and thigh injuries were significantly associated with greater hospitalization costs. CONCLUSIONS: Using quantile regression to identify factors associated with hospitalization costs could be helpful for addressing the burden of injury in the elderly population. Policymakers may find these findings to be insightful in lowering hospitalization costs related to injury in the elderly population.

Digital measurement and correlation analysis of coronal and sagittal anatomic parameters in the radiographs of adolescent patients with idiopathic scoliosis.

OBJECTIVE: To use digital software to measure the morphologic and anatomic parameters of adolescent idiopathic scoliosis (AIS). Differences and correlations among different parameters were compared to provide an anatomic basis for the selection of treatment methods and preoperative evaluation of AIS. METHODS: Spinal radiographs were taken from 300 boys and girls (age, 10-18 years) suffering from idiopathic scoliosis in four grade-A hospitals in Inner Mongolia. After screening, 120 cases with complete imaging data were assessed. Imaging data were transferred to a work station (Dr Wise™). The anatomic indices of the Cobb Angle, CVA, AVT, TS, CA, CPT, CSI, FPT, CCA, TK, LL, SS, PT, and PI were measured. RESULTS: There were significant differences in AVT between different grades and types of scoliosis (F = 34.079, P = 0.000; χ2 = 23.379, P = 0.000). AVT was a protective factor, and the smaller the AVT, the less severe was the scoliosis. Compared with adolescents with mild or moderate scoliosis, the Cobb angle of adolescents with severe scoliosis was negatively correlated with CCA, LL, and SS (r = - 0.641, p < 0.05; r = - 0.695, p < 0.01; r = - 0.814, p < 0.01). CONCLUSIONS: Some of the anatomic parameters in the coronal and sagittal planes of adolescents with idiopathic scoliosis were significantly different according to the severity and type of scoliosis. Significant correlations were found between more anatomic indices in adolescents with severe scoliosis than in adolescents suffering from mild or moderate scoliosis.

Daphnetin alleviates neuropathic pain in chronic constrictive injury rats via regulating the NF-κB dependent CXCL1/CXCR2 signaling pathway.

CONTEXT: Daphnetin is a natural product with anti-inflammatory, antioxidant, and neuroprotective properties. Reports have found that it has a strong analgesic effect; however, its analgesic mechanism is unknown. OBJECTIVE: We explored the effect and mechanism of daphnetin on neuropathic pain (NP). MATERIALS AND METHODS: The rat model of NP was established by ligation of the sciatic nerve. Male Sprague-Dawley rats were divided into six groups: Control, Model, Sham, morphine (0.375 mg/kg), and daphnetin (0.0625 and 0.025 mg/kg). Rats were intrathecally injected with drugs or normal saline once daily for three days. Hyperalgesia was evaluated by mechanical withdrawal threshold (MWT) and thermal withdrawal threshold (TWT). Protein levels were detected using ELISA, immunofluorescence, and western blotting. RESULTS: Compared to the Model group, daphnetin improved TWT (46.70 °C vs. 42.20 °C) and MWT (45.60 g vs. 23.60 g), reduced the expression of interleukin-1ß (0.99 ng/g vs. 1.42 ng/g), interleukin-6 (0.90 ng/g vs. 1.52 ng/g), and tumor necrosis factor-α (0.93 ng/g vs. 1.52 ng/g) in the sciatic nerve. Daphnetin decreased the expression of toll-like receptor 4 (TLR4) (0.47-fold), phosphorylated inhibitor of NF-κB (p-IKBα) (0.29-fold), nuclear factor kappaB (NF-κB) (0.48-fold), glial fibrillary acidic protein (GFAP) (0.42-fold), CXC chemokine ligand type 1 (CXCL1) (0.84-fold), CXC chemokine receptor type 2 (CXCR2) (0.78-fold) in the spinal cord. DISCUSSION AND CONCLUSIONS: Daphnetin alleviates NP by inhibiting inflammation and astrocyte activation in the spinal cord, providing theoretical support for the extensive clinical treatment of NP.

Association of variants in KCNK17 gene with ischemic stroke and cerebral hemorrhage in a Chinese population.

BACKGROUND: KCNK17 (potassium channel, subfamily K, member17) has a role in the pathogenesis of stroke. We reported previously that rs10947803 single-nucleotide polymorphism (SNP) in KCNK17 is associated with cerebral hemorrhage in a Chinese population. The aim of the present study was to examine other SNPs in the KCNK17 gene that are associated with cerebral hemorrhage and other subtypes of stroke in the Chinese population. METHODS: A total of 1356 subjects with stroke and 1225 control patients were examined by a case-control methodology. The SNPs (rs12214600, rs12195376, rs2758912, and rs10807204) in KCNK17 gene were genotyped with the TaqMan real-time polymerase chain reaction assay. RESULTS: rs12214600 SNP in KCNK17 was significantly associated with cerebral hemorrhage (unadjusted odds ratio = .55, 95% confidence interval = .35-.86, P = .008, q = .0328) under the allele model. After adjusting for age, sex, and hypertension, we found that the association remained significant (odds ratio = .56, 95% confidence interval = .35-.90, P = .0158). There was no association detected for other SNPs in KCNK17 with cerebral hemorrhage, and none of the SNPs in KCNK17 had an association with ischemic stroke. CONCLUSIONS: The T carrier of an SNP (rs12214600) is associated with reduced risk of cerebral hemorrhage in the Chinese population, together with previous findings that SNPs rs10947803 and rs12214600 in the KCNK17 gene are associated with hemorrhagic stroke, but none of the SNPs tested had an association with ischemic stroke. KCNK17 may be important in the pathogenesis of cerebral hemorrhage.

[A case-control study on the association between cigarette smoking and ischemic stroke].

OBJECTIVE: To investigate the association between cigarette smoking and ischemic stroke. METHODS: A case-control study was conducted. Investigators got the available information from the cases and controls by questionnaires. Logistic regression models were used to analyze the relationship between smoking and ischemic stroke in this study. RESULTS: A total number of 1037 cases and 1205 controls were included in this study. As the results showed, cigarette smoking was associated with ischemic stroke (OR = 1.368, 95% CI 1.158-1.616). After adjustment for age, body mass index, waist hip ratio, blood pressure, blood glucose, blood lipid, stroke family history and alcohol, the relationship between smoking and ischemic stroke was still significant (OR =2. 158, 95% CI 1.293-3.600). And the more you smoke, the more you stroke. Compared with nonsmokers, the ORs of smokers with 1-9 / day, 10-19 / day and 20 - / day were 1.097, 1.168 and 2.950. CONCLUSION: Cigarette smoking is a risk factor for ischemic stroke.

How heavy is the medical expense burden among the older adults and what are the contributing factors? A literature review and problem-based analysis.

In recent years, the aging population and increasing medical expenses among the older adults have emerged as significant public health concerns. National governments must conduct medical expense accounting and implement measures to reduce the burden of medical costs on the older population. However, limited studies have focused on total medical expenditure from a macro perspective, with many researches exploring individual medical expenses from different perspectives. This review introduces the trend of population aging and its impact on health cost change, reviews research on the medical expense burden of the older population and contributing factors, and points out underlying problems and limitations of current studies. Based on the present studies, the review emphasizes the necessity of medical expense accounting and analyzes the medical expense burden of the older population. Future studies should explore the impacts of medical insurance funds and health service system reforms on reducing medical expenses and developing a supporting medical insurance reform plan.

The core of maintaining neuropathic pain: Crosstalk between glial cells and neurons (neural cell crosstalk at spinal cord).

BACKGROUND: Neuropathic pain (NP) caused by the injury or dysfunction of the nervous system is a chronic pain state accompanied by hyperalgesia, and the available clinical treatment is relatively scarce. Hyperalgesia mediated by pro-inflammatory factors and chemokines plays an important role in the occurrence and maintenance of NP. DATA TREATMENT: Therefore, we conducted a systematic literature review of experimental NP (PubMed Medline), in order to find the mechanism of inducing central sensitization and explore the intervention methods of hyperalgesia caused by real or simulated injury. RESULT: In this review, we sorted out the activation pathways of microglia, astrocytes and neurons, and the process of crosstalk among them. It was found that in NP, the microglia P2X4 receptor is the key target, which can activate the mitogen-activated protein kinase pathway inward and then activate astrocytes and outwardly activate neuronal tropomyosin receptor kinase B receptor to activate neurons. At the same time, activated neurons continue to maintain the activation of astrocytes and microglia through chemokines on CXCL13/CXCR5 and CX3CL1/CX3CR1. This crosstalk process is the key to maintaining NP. CONCLUSION: We summarize the further research on crosstalk among neurons, microglia, and astrocytes in the central nervous system, elaborate the ways and connections of relevant crosstalk, and find potential crosstalk targets, which provides a reference for drug development and preclinical research.

Daphnetin Improves Neuropathic Pain by Inhibiting the Expression of Chemokines and Inflammatory Factors in the Spinal Cord and Interfering with Glial Cell Polarization.

Neuropathic pain (NP) is a common pain disease that seriously affects the quality of life and physical and mental health of patients. Daphnetin is extracted from the Daphne giraldii Nitsche and has the structure of 7,8-dihydroxy coumarin. As a natural product, daphnetin displays a wide range of pharmacological activities, such as analgesia and anti-inflammatory activities, but whether it is able to improve NP through anti-inflammatory effects is unknown. Therefore, this paper intends to investigate the mechanism of daphnetin in improving NP rats affected by the intrathecal injection of tumor necrosis factor-α (TNF-α) from the perspective of anti-inflammation. Our results showed that daphnetin significantly improved hyperalgesia in NP rats. Daphnetin inhibited the activation and polarization of glial cells and neurons in the spinal cord of NP rats and reduced the expression of mRNA and protein of inflammatory factors and chemokine pairs in the spinal cord. Daphnetin inhibited the polarization of human microglia cell 3 (HMC3) cells and human glioma cells (U251) cells toward M1 microglia and A1 astrocytes, respectively, and induced the conversion of M1 microglia and A1 astrocytes to M2 microglia and A2 astrocytes, respectively. In conclusion, daphnetin ameliorates NP by inhibiting the expression of inflammatory factors and chemokines and the polarization of glial cells in the spinal cord of NP rats. This study provides a theoretical basis for the treatment of NP with daphnetin to expand the clinical application of daphnetin.

Chinese Herbal Medicine Alleviates Myocardial Ischemia/Reperfusion Injury by Regulating Endoplasmic Reticulum Stress.

Myocardial ischemia/reperfusion injury is the main cause of increased mortality and disability in cardiovascular diseases. The injury involves many pathological processes, such as oxidative stress, calcium homeostasis imbalance, inflammation, and energy metabolism disorders, and these pathological stimuli can activate endoplasmic reticulum stress. In the early stage of ischemia, endoplasmic reticulum stress alleviates the injury as an adaptive survival response, but the long-term stress on endoplasmic reticulum amplifies oxidative stress, inflammation, and calcium overload to accelerate cell damage and apoptosis. Therefore, regulation of endoplasmic reticulum stress may be a mechanism to improve ischemia/reperfusion injury. Chinese herbal medicine has a long history of clinical application and unique advantages in the treatment of ischemic heart diseases. This review focuses on the effect of Chinese herbal medicine on myocardial ischemia/reperfusion injury from the perspective of regulation of endoplasmic reticulum stress.

[Study on characterization of the complexes of FUS1/hIL-12 with cationic liposome].

This study was aimed to shed light on the biological and pharmaceutical characterization of the complexes of FUS1/hIL-12 double gene with cationic liposome, and to assess such complexes' transfection efficiency, stability and cytotoxicity; for they have the potential for use as drugs in gene therapy of lung cancer. Gel retardation assay, diameter measurement, and surface charge by photon correlation spectroscopy (PCS) were employed to select the appropriate ratio of "cationic liposome to DNA" of the double-gene and liposome complexes. The plasmid EGFP and plasmid PVITO2-hIL12-FUS1 mediated by cationic liposome were transfected into A549 lung cancer cells respectively, and the expression levels of EGFP and FUS1 and hIL-12 were determined by inverted fluorescence microscope and immunohistochemical and enzyme linked immunosorbent assay (ELISA) respectively. Agarose gel electrophoresis was performed to detect the stability of the double-gene and liposome complexes, after they were incubated with serum and Dnase I respectively. After the erythrocytes being incubated with the complexes of FUS1/hIL-12 with cationic liposome, the morphology of erythrocyte was observed by microscopy. The result of this study provides a basis for the use of the complexes of FUS1/hIL-12 with cationic liposome in gene therapy of lung cancer.

[Impact the variability of blood pressure on the outcome of ischemic stroke patients upon discharge from the hospital].

OBJECTIVE: To explore the relationship between blood pressure variability and worsening functional outcomes of patients upon discharge from the hospitals. METHODS: The study cohort consisted of 404 patients who presented to Second Affiliated Hospital of Harbin Medical University with ischemic stroke during March 2012-March 2013. Systolic BP and diastolic BP were measured for each patient from admission to the fifth day and coefficient of variation blood pressure calculated. Disability at discharge was measured by the modified Rankin score (mRs). Chi-square test, t-test and multivariate logistic regression analysis were performed. RESULTS: After adjustment for potential confounding factors including age, sex, activity, smoking, alcohol intake,BMI, heart rate, hypertension, diabetes mellitus, stroke history, lipid parameters, homocysteine and FPG, results from the multivariate logistic regression analysis showed that when DBP variability was greater than 9, it was associated with a significantly worse functional outcome at hospital discharge compared with those less than 9, with the odds ratio as 1.70 (95%CI:1.02-2.84). When comparing the ones that DBP variability more than 10 with the ones less than 10, the odds ratio was 1.86 (95% CI:1.11-3.13). However, there was no significant association seen between SBP variability and the worse functional outcome at hospital discharge. CONCLUSION: Blood pressure variability might be associated with ischemic stroke outcome at hospital discharge but needed more evidence to approve.

Association of CVD candidate gene polymorphisms with ischemic stroke and cerebral hemorrhage in Chinese individuals.

BACKGROUND: Contribution of cardiovascular disease related genetic risk factors for stroke are not clearly defined. We performed a genetic association study to assess the association of 56 previously characterized gene variants in 34 candidate genes from cardiovascular disease related biological pathways with ischemic stroke and cerebral hemorrhage in a Chinese population. METHODS: There were 1280 stroke patients (1101 with ischemic stroke and 179 with cerebral hemorrhage) and 1380 controls in the study. The genotypes for 56 polymorphisms of 34 candidate genes were determined by the immobilized probe approach and the associations of gene polymorphisms with ischemic stroke and cerebral hemorrhage were performed by logistic regression under an allelic model. RESULTS: After adjusting for age, sex, BMI and hypertension status by logistic regression analysis, we found that NPPA rs5063 was significantly associated with both ischemic stroke (odds ratio [OR] 0.69; 95% confidence interval [CI], 0.52 to 0.90; P = 0.006) and cerebral hemorrhage(OR = 0.39; 95%CI, 0.19 to 0.78; P = 0.007). In addition, MTHFR rs1801133 also was associated with cerebral hemorrhage (OR = 1.48; 95%CI, 1.16 to 1.89; P = 0.001) but not with ischemic stroke (OR = 1.08; 95%CI, 0.96 to 1.22; P = 0.210). After false discovery rate (FDR) correction, the association of NPPA rs5063 and MTHFR rs1801133 with cerebral hemorrhage remained significant. CONCLUSIONS: The NPPA rs5063 is associated with reduced risk for cerebral hemorrhage and MTHFR rs1801133 is associated with increased risk of cerebral hemorrhage in a Chinese population.

Synergistic effects of eukaryotic coexpression plasmid carrying LKB1 and FUS1 genes on lung cancer in vitro and in vivo.

PURPOSE: LKB1 and FUS1 are two kinds of new tumor suppressor genes as well as early-stage genes in lung cancer. Recent studies showed that LKB1 and FUS1 play important roles in lung carcinogenesis process. We hypothesized that combined gene therapy with LKB1 and FUS1 could inhibit lung cancer growth and development synergistically. METHODS: In this study, two kinds of tumor suppressor genes, LKB1 and FUS1, were constructed in an eukaryotic coexpression plasmid pVITRO(2), and then, we evaluated the synergistic effects of the two genes on anticancer activity and explored the relevant molecular mechanisms. RESULTS: We defined coexpression of LKB1 and FUS1 could synergistically inhibited lung cancer cells growth,invasion and migration and induced the cell apoptosis and arrested cell cycle in vitro. Intratumoral administration of liposomes: pVITRO(2)­LKB1­FUS1 complex (LPs­pVITRO(2)­LKB1­FUS1) into subcutaneous lung tumor xenograft resulted in more significant inhibition of tumor growth. Furthermore, intravenous injection of LPs­pVITRO(2)­LKB1­FUS1 into mice bearing experimental A549 lung metastasis demonstrated synergistic decrease in the number of metastatic tumor nodules. Finally, combined treatment with LKB1 and FUS1 prolonged overall survival in lung tumor-bearing mice. Further study showed tha tthe synergistic anti-lung cancer effects of coexpression ofLKB1 and FUS1 might be related to upregulation of p-p53, p-AMPK and downregulation of p-mTOR, p-FAK, MMPs, NEDD9, VEGF/R and PDGF/R. CONCLUSIONS: Our results suggest that combined therapy with eukaryotic coexpression plasmid carrying LKB1 and FUS1 genes may be a novel and efficient treatment strategy for human lung cancer.

[A case-control study on the duration of sleeping and cerebral infarction].

OBJECTIVE: To explore the relationship between duration of sleeping and cerebral infarction. METHODS: A case-control study involved 1037 cerebral infarction patients admitted by the Second Affiliated Hospital of Harbin Medical University,December 2011-December 2012 as cases. Another 1205 adults free from cerebro-vascular diseases who had undergone physical examination in the hospital at the same period, were served as controls. All the subjects were interviewed with unified questionnaire. Chi-square test, u-test and multivariate logistic regression analysis were performed. RESULTS: After adjustment for potential confounding factors including age, sex, body mass index, wrist-hip ratio, smoking, alcohol intake, hypertension, diabetes mellitus, coronary artery disease and lipid parameters, data from the multivariate logistic regression analysis showed that the risk of cerebral infarction was greater in people who slept less than 6 hours per night than those who slept between 6 hours and 8 hours per night, with an odds ratio (95% CI)as 2.81 (95% CI:1.68-4.70). There was no significant association between factor as 'sleeping longer than 8 hours/pre day' and cerebral infarction. Through the subgroup analysis, data showed that the association between 'shorter than 6 hour sleep/night' and cerebral infarction consistently existed, across the categories of sex, and the degree of association was greater in women than in men, with the odds ratio as 5.58 (95% CI: 1.78-17.52) and 2.00 (95% CI:1.10-3.64) respectively. CONCLUSION: Short sleeping duration might increase the risk of developing cerebral infarction.

Cationic liposome-mediated nitric oxide synthase gene therapy enhances the antitumor effects of cisplatin in lung cancer.

Cisplatin is one of the most effective antitumor drugs for non-small cell lung carcinoma (NSCLC) patients. However, its efficacy has encountered a plateau due to its side effects and drug resistance. Inducible nitric oxide (NO) synthase (iNOS) gene therapy has been reported to have antitumor effects in several types of cancers and enhances sensitivity to cisplatin, but the effects of iNOS gene therapy alone or its combination with cisplatin in lung cancer remain unclear. In the current study, we evaluated the effects of cationic liposome (LP)-mediated iNOS gene transfection on enhancing low-dose cisplatin-mediated antitumor effects in the A549 human lung adenocarcinoma cell line in vitro. Furthermore, we examined whether iNOS gene therapy enhances the antitumor effects of low-dose cisplatin in two A549 human lung cancer cell xenograft mouse models. The results revealed that iNOS gene therapy may significantly enhance low-dose cisplatin-mediated inhibition of cell proliferation, invasion, migration and promotion of cell apoptosis in A549 cells. Intratumoral administration of the LP-pVAX-iNOS complex significantly enhanced low-dose cisplatin-mediated suppression of subcutaneous tumor growth. Moreover, intravenous injection of the LP-pVAX-iNOS complex greatly enhanced low-dose cisplatin-mediated inhibition of experimental lung metastasis and prolonged the life span of mice without significant organ-related toxicity in a nude mouse model of lung metastasis compared to the cisplatin alone-treated group. Furthermore, iNOS gene-mediated enhancement of cisplatin-mediated antitumor effects in lung cancer may be related to the attenuation of p-mTOR, MMP2 and the activation of p-p53. Thus, the combination treatment with iNOS gene therapy and cisplatin may be a novel and effective therapeutic strategy for lung cancer.

[Construction of A eukaryotic expression vector carrying the iNOS gene and its effect on A549 lung cancer cells].

BACKGROUND AND OBJECTIVE: The iNOS gene is associated with NO-mediated antitumor effects. The aims of this study are to construct a eukaryotic expression plasmid that carries the iNOS gene and to detect the expression levels and antitumor effects of the iNOS gene on A549 lung cancer cells. METHODS: A DNA fragment of the human iNOS coding sequence was amplified using reverse transcription polymerase chain reaction (RT-PCR). The DNA fragment was subsequently cloned into the multiple cloning sites of the eukaryotic expression vector pVAX. The recombinant plasmid was confirmed using restriction enzyme treatment, PCR, and sequencing and was then transfected into A549 lung cancer cells. The expression of the iNOS gene in the A549 lung cancer cells after transfection was verified by RT-PCR and Western blot analysis. The effects of iNOS on cell apoptosis, proliferation, and migration were identified by staining with Hoechst 3235, an MTT assay, and a scratch assay, respectively. RESULTS: The results of the restriction enzyme digestion, PCR, and sequencing verified the successful construction of the eukaryotic expression plasmid pVAX-iNOS. The iNOS gene expression level was increased in the transfected A549 cells. Further experiments also showed increased cell apoptosis among the A549 lung cancer cells transfected with pVAX-iNOS. Meanwhile, the proliferation and migration of A549 cells were significantly inhibited by the enhanced iNOS gene expression. CONCLUSION: The recombinant eukaryotic expression vector pVAX-iNOS was successfully constructed and transfected into A549 cells. The enhanced iNOS gene expression significantly promoted cell apoptosis, whereas the proliferation and migration of A549 cells were inhibited. These findings contribute to the development of novel and effective gene therapies for lung cancer.

Cationic lipids containing cyclen and ammonium moieties as gene delivery vectors.

In this study, two novel cationic lipids containing protonated cyclen and quaternary ammonium moieties were designed and synthesized as non-viral gene delivery vectors. The structures of the two lipids differ in their hydrophobic region (cholesterol or diosgenin). Cationic liposomes were easily prepared from the lipids individually or from the mixtures of each cationic lipid and dioleoylphosphatidylethanolamine. Several studies including DLS, gel retardation assay, and ethidium bromide intercalation assay suggest that these amphiphilic molecules are able to bind and compact DNA into nanometer particles which can be used as non-viral gene delivery agents. Our results from in vitro transfection show that in association with dioleoylphosphatidylethanolamine, two cationic lipids can induce effective gene transfection in human embryonic kidney 293 cells, although the gene transfection efficiencies of two cationic lipids were found to be lower than that of lipofectamine 2000(TM) . Besides, different cytotoxicity was found for two lipoplexes. This study demonstrates that the title cationic lipids have large potential to be efficient non-viral gene vectors.

Novel cationic lipids possessing protonated cyclen and imidazolium salt for gene delivery.

In this study, two novel protonated cyclen and imidazolium salt-containing cationic lipids differing only in their hydrophobic region (cholesterol or diosgenin) have been designed and synthesized for gene delivery. Cationic liposomes were easily prepared from each of these lipids individually or from the mixtures of each cationic lipid and dioleoylphosphatidyl ethanolamine (DOPE). Several studies including DLS, gel retardation assay, ethidium bromide intercalation assay, and TEM demonstrated that these amphiphilic molecules are able to bind and compact DNA into nanometer particles that could be used as non-viral gene delivery agents. Our results from in vitro transfection showed that in association with DOPE, two cationic lipids can induce effective gene transfection in HEK293 cells. Furthermore, the gene transfection efficiencies of two cationic lipids were dramatically increased in the presence of calcium ion (Ca(2+)). It is notable that the gene transfection abilities of two cationic lipids were maintained in the presence of 10% serum. Besides, different cytotoxicity was found for two lipoplexes. This study demonstrates that the title cationic lipids have large potential to be efficient non-viral gene vector.

Effect of Fraxiparine, a type of low molecular weight heparin, on the invasion and metastasis of lung adenocarcinoma A549 cells.

Lung cancer is one of the most highly malignant tumors, and a significant threat to human health. Lung cancer patients often exhibit tumor cell invasion and metastasis, which often render current treatments ineffective. Recently, the beneficial effects of low molecular weight heparin (LMWH) on cancer metastasis were reported in pre-clinical research studies. LMWH may be a potential drug for cancer therapy. However, the mechanism of LMWH on the invasion and metastasis of cancer has yet to be determined. This study investigated the effects of Fraxiparine on the proliferation, invasion and metastasis of the human lung adenocarcinoma A549 cell line. MTT assay and flow cytometry showed that Fraxiparine slightly inhibited the cell viability dose- and time-dependently, but did not arrest the A549 cells in the G1 phase nor induce early apoptosis. The transwell chamber assay showed that Fraxiparine significantly suppressed the invasion and migration of the A549 cells in vitro. Fraxiparine also markedly inhibited the adhesion of the A549 cells to Matrigel. The RT-PCR assay demonstrated that the reduction in invasion and metastasis may be related to the up-regulation of nm23-H1 and the down-regulation of the heparanase expression. Moreover, the RT-PCR assay and Western blot analysis demonstrated that down-regulation of the expression of integrin ß1 and ß3, as well as that of matrix metalloproteinase-2 and -9 may be responsible for the inhibition of the invasion and metastasis of A549 cells by Fraxiparine.