Objective:To observe the effects of salmeterol and fluticasone combined with the conventional treatment on the airway remodeling and bone metabolism in the patients with severe chronic obstructive lung disease ( chronic obstructive pulmonary disease, COPD. Methods:Totally 90 patients with severe COPD were divided into the observation group and the control group according to the order of admission. The control group was received the conventional treatment, and the observation group was treated with salmeterol and fluticasone additionally. The course of treatment was 8 weeks. Before the treatment and in the 3rd and 12th month after the treat-ment, the changes in the levels of bone mineral density ( BMD) in femoral neck bone, serum osteocalcin ( BGP) , alkaline phosphatase (AKP), the total number of induced sputum cells, neutrophils percentage, forced expiratory volume in one second (FEV1), leukocyte mediated element 8 (IL-8), matrix metalloproteinase 9 (MMP-9) and other indicators were detected and compared. Results: BGP, AKP and BMD of the two groups in the 3rd month after the treatment were not significantly changed (P>0. 05), while in the 12th month after the treatment, BMD in the observation group was significantly lower than that in the control group and that before the treat-ment, and BGP and AKP in the observation group were significantly higher than those in the control group and that before the treatment (P<0. 05). The total number of induced sputum cells, the percentage of neutrophils, FEV1, MMP-P and IL-8 in the two groups were significantly improved in the 12th month after the treatment (P<0. 05), and the difference between the two groups was statistically sig-nificant except IL-8 (P<0. 05). Conclusion:Salmeterol and fluticasone propionate can effectively improve airway remodeling in the patients with COPD. Short time use of salmeterol and fluticasone propionate shows no adverse effect on the bone metabolism, while long-term use would lead to reduced bone mineral density.

AIM To study in vitro stereoselective metabolism of mandelic acid (MA) in 3 kinds of laboratory animal tissue fractions and observe the differences of MA biotransformation. METHODS MA enantiomers were incubated with the tissue fractions from rats, mice and rabbits. The phenylglyoxylic acid (PGA) concentrations in incubation mixture were determined by high-performance liquid chromatography. Coenzymes and inhibitors were co-incubated with MA to investigate their effects on MA metabolism. RESULTS Only S-MA showed a unique metabolism in liver and kidneys S9 fractions of rats. No inhibition of the enzyme activity was observed by addition of ethanol or 4-methyl pyrazole. NADPH caused a remarkable increase in S-MA metabolism. CONCLUSION Nonmicrosomal enzymes in kidneys and liver except alcohol dehydrogenase are responsible for the stereoselective metabolism of MA in rats. The MA biotransformation is significantly different between rats and mice or rabbits.

Objective To investigate the vascular endothelial growth factor (VEGF) expression level by chondrocytes isolated from patients with osteoarthritis (OA) in hip or femoral neck fracture (FNF) and explore the effect of synovial fluid from OA or FNF on secretion of VEGF. Methods The cartilage tissues were collected from 12 patients with OA in hip and 8 patients with FNF. Cartilage was stained with HIM and Safranin O/Fast Green (S/F) method. The damage of cartilage was evaluated using Mankin scores.Cathepsin B which was selected for cell dedifferentiation monitoring marker and VEGF level was detected in the supernatant fluid. The synovial fluid from OA, FNF and DMEM were respectively added to the culture medium to explore their effects on regulating VEGF. Results Cartilage the Mankin scores of OA group were higher than that of FNF group. Chondrocytes gradually lost their original spherical appearance, with Cathepsin B upregulated while VEGF downregulated. The OA synovial fluid can stimulate chongdrocytes to secrete more VEGF than the one from patients with FNF. However, chondrocytes gradually produced less VEGF after passaging. Conclusion Mankin scores had good correlation with chondrocytes' VEGF production in the early stage of primary culture. Chondrocytes showed quick dedifferentiation characteristics in vitro. OA synovial fluid showed abig ger capability in stimulating chondrocytes to express more VEGF, which might indicate that OA synovial fluid participated in the pathological process of OA.

The goal of treatment of metabolic syndrome is the prevention of diabetes and cardiovascular events. A series of novel tetrahydrocoptisine quaternary ammonium compounds were prepared to evaluate their action of hypoglycemia and hypolipidemia for finding the therapeutic agents of metabolic syndrome. Starting from the coptisine hydrochloride (2), fifteen target compounds were synthesized by reduction and substitution of the 7-N position. All of the target compounds were characterized by 1H NMR and HR-MS. Their hypoglycemic activities were evaluated in HepG2 cell and hypolipidemic activities of compounds with better hypoglycemic activity were tested further in vivo. Results indicated that compounds 5, 7, 8 and 9 exhibited better hypoglycemic activities in vitro and compounds 5 and 8 exhibited good hypolipidemic activities in high-fat-diet (HFD) induced hyperlipidemia mice and (or) hamsters. However, the activity is not as good as simvastatin.

Objective To compare the technical success ,complication rates and comfort of the radiologic placement of central venous ports(CVP) via the internal jugular vein ,subclavian vein via subclavian region ,subclavian vein via supraclavicular region un‐der DSA guidance .Methods We retrospectively reviewed 188 CVP patients implanted at hospitals between December 2012 and De‐cember 2013 .The patients were divided into three groups according to the different catheter implantation sites ,internal jugular vein (group A) ,subclavian vein via subclavian region (group B) and subclavian vein via supraclavicular region (group C) .Intraoperative pain score ,technical success rates ,peri‐procedural ,as well as early and late complication rates were recorded based on the image fol‐low‐up and patient medical records .Results The technical success rate was 100 .0% without any lethal complications .The CVP re‐lated infections were occurred in each group with 1 patient ,but there was no statistics significant different(P>0 .05) .Subclavian vein thrombosis were occurred 2(2 .1% ,2/96) in group B ,which was recanalized after thrombolytic therapy ,and 1(1 .7% ,1/60) in group C ,which was completely occluded .The higher rate of catheter migration and kinking of catheter were occurred in group A and group C .The rate was statistically significant difference among the three groups (P<0 .01) .The late complication rates were statistically higher in group A and C compared with group B (P<0 .05) .The punch‐off was just occurred in group B (n=2) .There were no significant differences about catheter fracture ,port rotation and wound dehiscence among the three groups(P>0 .05) .Con‐clusion In DSA radiologic placement of a CVP via the subclavian vein via subclavian region is safe and efficient with more comfort‐able and lower complication rates ,which could be chosen priority .

Objective: To explore the characteristics and relationship between physical activity and self-efficacy of college students during the COVID-19 epidemic, so as to provide evidence for the orderly development of physical education and curriculum reform of college students after their return to school. Methods: A total of 6 227 college students were collected from 8 districts of China by using convenient sampling method and snowball sampling method, and the physical activity and self-efficacy of all participants were assessed used by the International Physical Activity Questionnaire Short Form (IPAQ-S) and the General Self-Efficacy Scale(GSES). Results: In the terms of general self-efficacy, the score of male students was significantly higher than that of female students, the score of freshman was significantly higher than that of other grades(t/F=7.32, 5.56, P<0.01). In terms of physical activity, the middle and high level physical activity of girls was significantly higher than that of boys, but the proportion of boys meeting the recommended amount was higher(P<0.01). Low level physical activity was the highest in junior college students(31.75%), medium level physical activity was the highest in freshmen (46.75%) and sophomore student (48.13%), and high level physical activity was the highest senior college students (30.40%). The moderate level of physical activity was the highest in all BMI groups, while the lean group had the lowest level of low level physical activity (15.70%) and the highest level of high level physical activity(37.31%), and the lowest proportion meeting the recommended amount(P<0.05). Self-efficacy had a positive predictive effect on physical activity(OR=1.04, 95%CI=1.03-1.05). Conclusion During the COVID-19 epidemic, there are significant differences in general self-efficacy and physical activity among college students by gender, grade and BMI. The general self-efficacy has a positive impact on physical activity, suggesting that college physical education should pay attention to students- self-efficacy, while considering individual and group differences, and promote physical activity level through self-efficacy improvement.

BACKGROUNDTo study the effects of extraneous p53 antisense RNA on malignant growth and sensitivity to cisplatin of human lung cancer cell line.

METHODS801D cell line with p53 deletion and mutation at 248 codon was selected as a parent cell line. An 1.8 kb human p53 full length cDNA was inserted into a mammalian expression vector PEGFP to construct a p53 antisense RNA recombined plasmid PEGFP-p53(AS) and GFP gene at plasmid was a report gene to monitor extraneous gene expression. The extraneous gene was detected by PCR. The p53 mutation protein was examined by immunohitochemical stain of p53 monoclonal antibody. The inhibition growth efficacy of extraneous p53 in vitro was determined by clonogenic survival assay. Sensitivity of cells to cisplatin was examined with MTT assay. FCM analysis was performed to measure the effect of p53 antisense RNA on cell cycle.

RESULTSTwo cell lines, PEGFP-p53(AS)-801D and PEGFP-801D, were established after transfection of 801-D cells by lipofection and selection. Presence of extraneous p53 gene in PEGFP-p53(AS)-801D was proved by PCR and expression of extraneous p53 was estimated when green fluorescence in those cells was found out under the fluorescent microscopy. Mutated p53 protein in parent cell line 801D was positive and in PEGFP-p53(AS)-801D was negative with immunochemical stain. The inhibition rate of colony formation was 61% for PEGFP-p53(AS)-801D (P < 0.001). The sensitivity of PEGFP-p53(AS)-801D cells to cisplatin was increased. FCM analysis showed that the cell line was arrested at G1 phase.

CONCLUSIONSp53 mutation at 248 code plays an important role on malignant growth and resistance to cisplatin of human lung cancer cell line 801D. Malignant growth of cells with p53 deletion and mutation at 248 codon can be inhibited by extraneous p53 antisense RNA, and simultaneously the sensitivity to cisplatin is also increased.

BACKGROUND: In the treatment of osteoporotic vertebral compression fractures with percutaneous vertebroplasty, the efficacy and safety of bone cement injection by unipedicular and bipedicular approaches are still controversial. Some studies suggest that bone cement injection via unipedicular approach can shorten operation time and reduce postoperative complications, while the other studies suggest that bone cement injection via bipedicular approach can make bone cement distribute more evenly in the vertebral body and relieve pain better. OBJECTIVE: To systematically assess the efficacy and safety of percutaneous vertebroplasty via unipedicular versus bipedicular approach in the treatment of osteoporotic vertebral compression fractures. METHODS: Randomized controlled trials about unipedicular versus bipedicular percutaneous vertebroplasty in the treatment of osteoporotic vertebral compression fracture published before September 18 t h, 2018 were retrieved in the PubMed, Cochrane library, Embase, CNKI, VIP, WanFang data and CBM. Two researchers independently screened all the literatures, carried out data extraction and used improved Jadad to evaluate the methodological quality of the included studies. Meta-analysis using Revam 5.3 was conducted. Egger's test was utilized to evaluate the publication bias. RESULTS AND CONCLUSION: Totally 14 randomized controlled trials including 900 cases were eventually included, 452 cases in unipedicular approach group and 448 cases in bipedicular approach group. The Meta-analysis results showed that compared with the bipedicular approach, the unipedicular approach required shorter operation time [weighted mean difference (WMD) =-16.59, 95％ confidence interval (CI) (-19.25, -13.94), P < 0.001], smaller amount of bone cement injected [WMD=-1.27, 95％ CI (-1.64, -0.89), P < 0.001], and had lower incidence of cement leakage [relative risk=0.70, 95％ CI (0.53, 0.92), P =0.01]. There were no significant differences in short-and long-term Visual Analogue Scale scores, short-and long-term Oswestry Disability Index scores, and the postoperative incidence of adjacent vertebral fractures between the two groups (P> 0.05). Overall, bone cement injection both via bipedicular and unipedicular approaches can lead to a significant improvement in pain relief and living quality of osteoporotic vertebral compression fracture patients, but bone cement injection via unipedicular approach can shorten operation time, reduce cement volume and lower the incidence of cement leakage compared with the bipedicular approach.

Objective:To elucidate the effect of P2X4 signal axis on iron metabolism in the substantia nigra (SN) of male rats with Parkinson′s disease (PD) successfully induced by 6-hydroxydopamine (6-OHDA).Methods:A total of 120 male rats were randomly divided into control group, 6-OHDA group (PD group), P2X4-gene virus (P2X4-positive intervention, P2X4-PI) group, P2X4-gene unloaded virus (P2X4-negative control, P2X4-NC) group, P2X4-PI+6-OHDA group (inject P2X4 gene virus first, then 6-OHDA two weeks later) and P2X4-NC+6-OHDA group (inject no-load gene virus first, then two weeks later with 6-OHDA) using a completely random numbers method, with 20 rats in each group. Brain stereotactic instrument was used to inject the corresponding grouped drugs into the left SN of rats. A behavioral test was performed two weeks after the modeling was completed to select the qualified rat models, and the initiation and balance ability of each group of rats were further evaluated by a balance bar experiment. The brains of the qualified rat models were decapitated, and the brain tissue was taken away and preserved after related treatment. Immunofluorescence staining and Western blotting methods were used to detect the number of tyrosine hydroxylase (TH) positive dopaminergic neurons, and the expression levels of protein in P2X4 purinergic receptor (P2X4R), divalent metal-ion transporter-1 (DMT1) and ferroportin 1 (FPN1).Results:The results of immunofluorescence staining showed that the number of TH positive dopaminergic neurons in the left SN of the PD group (4 724.0±261.1, t=13.17, P<0.01) and the P2X4-NC+6-OHDA group (4 470.0±228.9, t=14.21, P<0.001) was significantly lower than that of the control group (7 942.0±461.6). The number of TH positive dopaminergic neurons of the P2X4-PI+6-OHDA group (2 493.0±371.6, t=8.092, P<0.01) was significantly lower than that of the P2X4-NC+6-OHDA group. The results of Western blotting suggested that compared with the control group (1.723±0.146, 1.369±0.107, 1.020±0.059), the expression of P2X4R, DMT1 was increased, whereas FPN1 was decreased in the left SN of the PD group (2.107±0.070, t=4.368, P<0.05; 1.733±0.117, t=4.245, P<0.05; 0.783±0.042, t=5.795, P<0.01) and the P2X4-NC+6-OHDA group (2.104±0.110, t=4.326; 1.737±0.073, t=4.291; 0.804±0.037, t=5.282; P<0.05). Compared with the P2X4-NC+6-OHDA group, the expression of P2X4R, DMT1 was increased and FPN1 was decreased in the left SN of the P2X4-PI+6-OHDA group (2.875±0.170, t=8.770; 2.845±0.180, t=12.92; 0.550±0.040, t=6.216; P<0.01). Conclusion:The overexpression of P2X4 gene can up-regulate the expression of DMT1 and down-regulate the expression of FPN1 in the SN, which leads to the deposition of iron in the SN of the midbrain, and then may cause damage to dopaminergic neurons, and finally has an effect on the occurrence and development of PD.

BACKGROUNDTo study the inhibition effects of both extraneous right sense and antisense p53 on malignant phenotype of human lung cancer cell line.

METHODSThe named 801D cell line with p53 deletion and mutation at 248 code was selected as a model in vitro. The recombined plasmid pEGFP-p53(RS) and pEGFP-p53(AS) were constructed. The extraneous gene was detected by PCR. The p53 mutation protein was examined by immunohistochemical stain of p53 antibody. The inhibition effect of extraneous p53 on tumor growth in vitro were determined by clonogenic survival assay. FCM analysis was carried out in cells. The inhibition effect on malignant growth of extraneous p53 in vivo was observed by heteroplastic transplant on nude mouse.

RESULTSThe transfected cell lines, pEGFP-p53(AS)-801D, pEGFP-p53(RS)-801D and pEGFP-801D were established. Presence of extraneous p53 and neo genes in pEGFP-p53(AS)-801D and pEGFP-p53(RS)-801D was proved by PCR and green fluorescence was found out in those cells under the microscope. Mutant protein in pEGFP-p53(AS)-801D was negative by immunohistochemical stain. The malignant growth of these transfected cell lines was inhibited comparing with parents in vivo and in vitro. Inhibition rate of colony formation was 62.0% for pEGFP-p53(AS)-801D and 80.8% for pEGFP-p53(RS)-801D. The tumorigenicity in nude mice was suppressed. Inhibition effects of extraneous right sense p53 on malignant growth of 801D was more distinct. FCM analysis showed that pEGFP-p53(AS)-801D cells were arrested at G1 phase.

CONCLUSIONSThe transfected cell lines with extraneous right sense and antisense p53 appear that malignant growth can be inhibited in vivo and in vitro.