Objective To construct the eukaryotic expression vector of mouse 24p3 gene and express it in NIH3T3 fibroblast cell line and verify its biological function of delivering iron into cells. Methods 24p3cDNA was cloned by RT-PCR and the eukaryotic expression vector of mouse 24p3 gene was constructed by gene recombination technique and the recombinant plasmid was verified by restriction enzyme digestion analysis and sequenceing, then transfected into NIH3T3 cells using DOTAP transfection reagent. Then the positive clones were selected and amplified. The expression of 24p3 gene in the culture supernatant of positive clones was analyzed by ELISA and Western blotting and its biological activity of delivering iron into cells was tested by atomic absorption spectrometer. Results The eukaryotic expression vector was constructed successfully. The NIH3T3 cells that stably expressed 24p3 were screened out. 24p3/SIP24 protein could express in the culture supernatant of positive clones and deliver iron into NIH3T3 cells. Conclusion The 24p3 gene was expressed successfully in the transfected NIH3T3 cells with good biological activity, which laid the foundation for further studying its biological function and mechanism involved in the iron metabolism.

The key to clinical paths administration lies in the optimization of the key processes and sectors in medical service, which will achieve an integration that is based on patients and span departments to cover all medical service sectors and processes. The clinical path practice was introduced into the hospital in 2005, and the clinical paths have been put into practice for over 20,000 cases/occasions by the end of 2009, scoring satisfactory social and economic outcomes as of now. For the purposes of speeding up the clinical path practice for expected outcomes in optimizing key medical service processes, we have got the following eight experiences and lessons: (1) Build shared demands; (2)Establish a powerful leadership; (3) Develop and promote visions; (4) Eliminate resistance and pool the forces; (5) Keep on the efforts and focus on details; (6) Reward involvements; (7) Recruit and develop key operators within the hospital; (8) Systemize and supervise the experiences of implementation.Promotion of clinical paths is a renovation in management with its inherent rules. Adoption of appropriate technical strategies and cultural strategies will upgrade the quality and acceptance of clinical path implementation.

Objective To study the value of umbilical artery(UA)combined with fetal middle cerebral artery(MCA)for detecting fetal intrauterine hypoxia.Methods A total of 349 pregnant women undergoing childbirth in our hospital from February 2015 and February 2016 were divided into the observation group(fetal intrauterine hypoxia group)and control group(normal fetus group)according to the existence of fetal intrauterine hypoxia.Then the systolic peak velocity and end-diastolic velocity ratio(S/D),pulse index(PI),resistance index(RI),(S/D)MCA/(S/D)UA,PIMCA/PIUA,RIMCA/RIUA of UA and MCA were compared between the two groups.The sensitivity,specific degree and Youden index of each index for diagnosing fetal hypoxia were statistically analyzed.Results The S/D,PI and RI values of UA at 28-35+6 pregnant weeks and at 36-41 pregnant weeks in the observation group were higher than those in the control group(P＜0.05),while the S/D,PI,RI value of MCA and(S/D)MCA/(S/D)UA,PIMCA/PIUA and RIMCA/RIUA values were lower than those in the control group(P＜0.01).The resistance index corresponding ratios of MCA and UA for diagnosing fetal hypoxia had the highest Youden index.Conclusion The combined use of fetal UA and MCA resistance index ratios has higher clinical value for predicting fetus intrauterine hypoxia.

Objective To observe the clinical efficacy of Daidzein Capsules(DC) on female climacteric syndrome.Methods Sixty eight patients with climacteric syndrome were randomized into DCgroup(n=37) and Gengnian'an control group (n=31).The treatment group and the control group were orally administered DC 150 mg and Gengnian'an Tablet 4.8 g,three times a day,respectively.The course of treatment lasted three months.Results The total effective rate in DC group was 91.89 %,which was superior to that in the control group.Time of drug becoming effective was shorter in DC group than that in the control group.After treatment,the level of E2 was significantly increased and the content of LH decreased significantly in DC group.Conclusion DC is safe and effective for the treatment of female climacteric syndrome.

Objective To observe the effects of low-frequency pulsed electromagnetic fields (LFPEMFs) on microcirculation angiogenesis in the hindlimbs of diabetic rats with acute ischemia. Methods Models of acute hindlimb ischemia were established in 60 male Sprague-Dawley diabetic rats. The diabetes model was established using 60 mg/kg intraperitoneal injections of streptozotocin (STZ). Fasting blood glucose levels were greater than 300 mg/dL. The rats were randomly divided into experimental and control groups. The rats in the experimental group were exposed to low-frequency pulsed electromagnetic fields for 2 hours each day, while the control group was not given any treatment. Laser-Doppler perfusion was used to measure blood flow in the ischemic hindlimb on days 0,7, 14 and 28 after the operation. The immunofluorescence of rat endothelial cell antigen-1 ( RECA-1) was used to evaluate the changes in angiogenesis. The levels of vascular endothelial growth factor (VEGF) and fibroblast growth factor-2 (FGF-2) were determined by both Western blotting and ELISA, and VEGFR2 and FGFR1 levels in the ischemic skeletal muscle were determined by Western blotting on days 7, 14 and 28 after the operation. Results The average perfusion ratio was significantly greater in the experimental group at days 14 and 28 compared with the control group. RECA-1 density in the tissues had increased significantly in the experimental group at the 14th and 28th day. The same was observed for FGF-2 and its receptor, but there was no significant difference for VEGF or its receptor in either group. Conclusions LFPMEFs can promote angiogenesis in acute hindlimb ischemia of diabetic rats by up-regulating FGF-2. This suggests that LFPMEFs may be useful for preventing and treating lower limb ischemia in diabetic humans.

Objective To prepare the chitosan-potD nanoparticles and to evaluate its protective efficacy against pneumococcal nasopharyngeal colonization. Methods potD gene was amplificated from pneumococcal genome and was inserted into pVAX1 expression vectors to construct pVAX1-potD recombinant plasmid which was then transfected into 293T cell using LipofectAMINE 2000 to analyze transient potD gene expression in vitro by RT-PCR and Western blot. Chitosan-potD nanoparticles were freshly prepared by coacervation methods at each time and the characterizations of the nanoparticles were then evaluated. BALB/c mice were immunized with chitosan-potD, naked potD DNA or pVAX1 for 4 times at two-week intervals. Anti-PotD IgG, IgG1 and IgG2a levels in serum and IgA levels in nasal washes, bronchoalveolar lavage fluids (BALF) and middle ear lavages(MEL) were detected by indirect enzyme-linked immunosorbent assay (ELISA). IL-17A, IL-4 and IFN-γ levels in splenocytes were determined by double sandwich ELISA. Mice were intrannsally challenged with Streptococcus pneumoniae ATCC6303, and Pneumococci were recovered from the nasopharyngeal niche at the fifth day after challenge. Results potD gene was successfully amplificated by PCR and the sequence was confimed to be consistent with that in the Genbank. The pVAX1-potD recombinant plasmid was successfully constructed and was expressed in eukaryocytes in vitro. The mean size and zeta potential of chitosan-potD nanoparticles was 430 nm and + 20.5 mv, respectively. Chitosan-potD nanoparticles were not digested by DNase Ⅰ , while naked potD DNA was completely digested. The levels of antibodies inculding IgG, IgG1, IgG2a, IgA and cytokines including IL-17A, IL-4 and IFN-γ were significantly higher in mice immunized with chitosan-potD nanoparticles than mice with naked potD or pVAX1 ( P <0.05) only. More importantly, much less Pneumococci were recovered from mice immunized with chitosan-potD nanoparticles than the other groups(P <0.05). Conclusion Chitosan-potD nanoparticles significantly enhanced the immunogenicity and protection efficacy of DNA vaccines by intranasal immunization and could be used as a potential mucosal vaccine to prevent pneumococcal infection.

Objective To observe the changes in levels of plasma leukocyte cell-derived chemnotaxin 2 (LECT2) in patients with newly diagnosed type 2 diabetes (T2DM) complicated with non-alcoholic fatty liver disease (NAFLD),and to investigate the clinical significance.Methods A total of 137 subjects were enrolled in the study,including 50 patients with newly diagnosed T2DM complicated with NAFLD,47 patients with newly diagnosed T2DM with non-NAFLD and 40 healthy subjects.The level of plasma LECT2 was determined by ELISA.Anthropometric data and other biochemical indicators were measured in three groups.The body mass index (BMI) and waist-to-hip ratio (WHR) were calculated.Insulin resistance and pancreas β-cell function were determined by homeostasis model assessment (HOMA-IR,HOMA-％β).Results Plasma concentration of LECT2 in patients with newly diagnosed T2DM complicated with non-NAFLD was higher than that in type 2 diabetic patients with non-NAFLD [(32.95 ± 10.11 vs 29.08 ± 7.54) ng/mL,P ＜ 0.01].Plasma LECT2 levels in both groups were significantly higher than that in normal control group [(22.38 ± 4.40) ng/mL,P ＜ 0.01].Plasma LECT2 level was positively correlated with BMI,FPG,FINS,C peptide,HbA1c,GGT,TG and HOMA-IR,while negatively with HDL-C and HOMA-％ β (all P ＜ 0.01).Multivariate regression analysis showed that levels of BMI,FPG and HDL-C were important factors affecting plasma LECT2 level.Conclusions Plasma LECT2 concentration significantly elevates in patients with newly diagnosed T2DM complicated with non-NAFLD.Plasma level of LECT2 is closely correlated with insulin resistance and glycolipid metabolism.LECT2 may play an important role in the patho genesis of insulin resistance and T2DM.

Objective To cultivate,identify and sort bronchoalveolar stem cells(BASC)derived from normal adult mouse lung.Methods After enzymatic digestion of lung tissue with dispase and collagenase in combination,the Sca-1+ cells were isolated from the obtained pulmonary cells by magnetic cell sorting.These Sca-1+ cells were cultured in dishes coated with collagen and mouse fibroblast cell line Swiss-3T3 under a serum-free culture system for BASC,which were identified by the dual-color immunofluorescent staining clara cell specific antigen(CCA)and surfactant protein C(SP-C).Finally,these pure BASC were isolated by the flow cytometry.Results One lung of normal adult mouse could yield(1.6-1.8)?107 nucleated cells in this enzyme digestion procedure.The percentage of Sca-1+ cells we sorted from lung tissue was much higher than the unsorted [(87.3?5.9)% and(9.6?1.8)%,P

Objective: To explore the value of evaluation of the diaphragmatic motion in chronic obstructive pulmonary disease (COPD) patients with dynamic X-ray chest radiography under calm breathing. Methods: Dynamic X-ray chest radiographs were performed in 96 COPD patients (COPD group) and 50 healthy controls (control group) under calm breathing. The range, speed and time of the diaphragmtic motion were obtained and analyzed. Results of lung function tests of 2 groups were recorded. Dynamic X-ray chest radiographs and pulmonary function parameters were compared between 2 groups, and the correlations of diaphragmatic motion parameters and lung function parameters were analyzed, respectively. Results: The left and right side diaphragmatic motion amplitude in COPD group ([18.63±7.45]mm, [15.64±5.98]mm) were larger than those in control group ([14.69±5.40]mm, [13.13±4.84]mm, both P<0.05). During inspiratory phase and expiratory phase, the motion speeds of left diaphragm in COPD group were 18.79 (15.67, 22.67) mm/s and 16.00 (12.36, 21.23) mm/s, the right side were 17.00 (12.71, 19.73) mm/s and 13.75 (9.50, 16.83) mm/s, respectively, while in control group of the left side were 15.33 (13.96, 17.71) mm/s and 11.42 (9.63, 16.00) mm/s, of the right side were (14.91±4.38) mm/s and (10.66±3.66) mm/s, respectively (all P<0.05). There were statistical differences of lung function between 2 groups (all P<0.01), but the pulmonary function parameters were not significantly correlated with dynamic chest radiograph examination parameters in COPD group (all P>0.05). Conclusion: Using dynamic X-ray chest radiograph could quantitatively analyze the diaphragmatic movement of COPD patients, providing a new examination method for accurate clinical evaluation of COPD.

Objective: To evaluate the efficacy of artificial intelligence (AI)-assisted pulmonary nodule diagnosis system in predicting the malignant probability of pulmonary subsolid nodule (SN). Methods: Pulmonary SN from 86 patients who underwent surgical resection for pulmonary space-occupying lesions were enrolled and divided into 3 groups according to post operation pathological results, i.e. preinvasive lesions (including atypical adenomatous hyperplasia[AAH]and adenocarcinoma in situ [AIS]) in group 1, microinvasive adenocarcinoma in group 2 and invasive adenocarcinoma in group 3, respectively. Preoperative chest CT data were imported into AI pulmonary nodule diagnosis system to measure CT value and volume, also malignant probability prediction value of each SN. The differences of volume, CT value and malignant probability of SN based on plain and enhanced CT were compared among 3 groups, while the volume, CT value and malignant probability of SN were compared between plain CT and enhanced CT in each group, respectively. The correlations of the predicted malignant probability of all SN according to 3 phase CT images and nodule density and volume were analyzed, respectively. Results: A total of 88 SN were enrolled, including 27 in group 1, 28 in group 2 and 33 in group 3. The sensitivity of all SN detected by AI system was 100% (88/88). The malignant probability of SN based on plain CT, arterial phase and delayed phase of enhanced CT was (85.18 [56.64, 92.08])%, (67.15 [58.99,90.30])% and (89.82 [56.64, 92.23])% in group 1, (93.10 [85.72, 95.75])%, (89.61 [74.44,95.35])% and (92.21 [86.74, 95.59])% in group 2, (97.05 [92.81, 98.74])%, (96.89 [90.40, 98.60])% and (96.49 [89.89, 98.69])% in group 3, respectively. Statistical differences of nodule volume, CT value and the malignant probability of 3 phases CT images were found among 3 groups (all P<0.01), while no statistically difference of malignant probability of SN between plain and enhanced CT was found in any group (all P>0.05). The nodule CT values of arterial phase and delayed phase in each group were significantly higher than that of plain CT (all P<0.01).The predicted malignant probabilities according to plain CT, arterial phase and delayed phase enhanced CT were all positively correlated with CT value and volume of SN (all P<0.01). Conclusion: The deep learning-based AI-assisted pulmonary nodule diagnosis system can assist in evaluation on the invasiveness of SN of pulmonary adenocarcinoma based on plain CT data, while enhanced CT has no significant effect on the efficiency of predicting malignant probability.