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For improving the corrosion resistance of carbon steel in the saline medium with high temperature and CO2-containing environment, tetrabutyl titanate was used to crosslink with urushiol under the compatibilization condition of propylene glycol methyl ether acetate to form a urushiol titanium coating, which was coated on carbon steel substrate surface for preparing high temperature resistant coating. Subsequently, thermogravimetry (TG) was employed to determine the thermal-resistant of urushiol titanium coating. The morphological and Fourier transform infrared spectroscopy (FTIR) were conducted to evaluate the protective capacity of the coating on carbon steel in simulated saline water by immersion test under high temperature, high pressure and CO2-containing medium. Moreover, electrochemical impedance spectroscopy (EIS) was utilized to evaluate the impedance change of coating in corrosive medium before and after immersion. Results showed that the thermal decomposition temperature of urushiol titanium coating achieved at 275.19 ℃. After immersed in simulated saline solution at 140 ℃ and 155 ℃ with CO2 pressure of 2 MPa for 30 d, the low-frequency impedance values of the coating could reach the respective magnitude of 1011 Ω·cm2 and 1010 Ω·cm2. Moreover, the valence bond characteristics of the coating did not change, and the surface morphology of coating was uniform and closely combined with the substrate, which indicated that the coating showed excellent protective performance against high temperature.

Starch-lipid composite nanoparticles were prepared by using dry and wet methods.The properties of nanoparticles,such as the lipid composite index,solubility,microscopic morphology,particle size distribution,three-phase contact angle,crystal structure,and emulsifying property,were studied to create a new starch-based Pickering emulsifier.The results show that the lipid composite index of the nanoparticles prepared by using the dry method is significantly higher than by using the wet method,which is related to the combination form between lipids and starch demonstrated by X-ray.Scanning electron microscopy results show that both particles exhibited the aggregated nanoscale distribution,and the average particle size of the dry method is significantly higher than the wet.The contact angle of the nanoparticles prepared with the dry method is significantly higher than that with the wet method.Emulsification property results show that the Pickering emulsion stabilized by nanoparticles prepared with the dry method has a smaller particle size and a more even distribution.Pickering emulsion gels were formed after 7 days of storage with the addition of 3%～5% emulsifier.

Objective: To investigate the effect of ginsenoside Rb1 (Gn-Rb1) on oxeiptosis in hepatocellular carcinoma (HCC) HepG2 cells and its possible molecular mechanism. Methods: Bioinformatic was used to analyse the effect of the expression of PGAM5, a key gene of oxeiptosis, on the survival time of HCC patients. HCC tissues and paracancerous tissues of 8 HCC patients admitted into Liaoning Provincial Tumor Hospital were selected, and the expressions of oxeiptosis-related gene proteins and mRNAs were detected by WB assay and qPCR. HepG2 cells were randomly divided into the control group and the Gn-Rb1 group (intervened with 200 µmol/L Gn-Rb1). The effects of Gn-Rb1 on the colony formation ability and the migration ability of HepG2 cells were detected by the cell clone formation assay and the cell scratch assay, respectively. ELISA was used to detect the effect on the level of ROS. Microplate was used to detect the effect on the level of LDH; the effects of Gn-Rb1 on the expressions of key gene proteins and mRNAs were detected by WB assay and qPCR. Results: Bioinformatic analysis revealed that, compared with low-expression patients, patients with PGAM5 high-expression hepatocellular carcinoma had longer overall survival time (P<0.05). In clinical hepatocellular carcinoma and paracancerous tissue samples, it was found that compared with paracancerous tissues, at protein and mRNA levels, the expressions of KEAP1 and PGAM5 in tumor tissues were significantly lower; the expression of NRF2 was significantly higher (all P<0.01); and the expression of p-AIFM1 was significantly higher (P<0.05). After HepG2 cells were intervened with 200 µmol/L Gn-Rb1, compared with those in the control group, the migratory and colony-forming abilities of HepG2 cells in the Gn-Rb1 group reduced significantly (all P<0.01), and the level of LDH increased significantly (P<0.05). Compared with the control group, at mRNA and protein levels, the expressions of KEAP1 and PGAM5 in the Gn-Rb1 group were significantly higher (all P<0.05); the expression of NRF2 was significantly lower (all P<0.05); and the expression of p-AIFM1 protein was significantly lower (P<0.01). Conclusion: Oxeiptosis is inhibited in hepatocellular carcinoma tissues, and Gn-Rb1 promotes oxeiptosis in HCC HepG2 cells by regulating the KEAP1/PGAM5/AIFM1 pathway, thus suppressing the proliferation and migration abilities of HepG2 cells. [ABSTRACT FROM AUTHOR]

AIM: Using bioinformatics analysis and experiment validation to explore the differential expression genes related to abnormal lipid metabolism in hepatocellular carcinoma (HCC) and the molecular mechanism of pachymaran affecting pyroptosis through squalene epoxidase (SQLE)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3)/gasdermin D (GSDMD) signaling pathway. METHODS: (1) The GEO, GSEA, DAVID, STRING and GEPIA databases were employed to screen abnormal lipid metabolism-related differentially expressed genes in HCC. (2) The tumor tissues from HCC patients (n=9) were collected to verify the differential expression of SQLE. (3) The inhibitory effect of pachymaran on the viability of human HCC cell line HepG2 was measured by CCK-8 assay. (4) The HepG2 cells were divided into control group and pachymaran (800 mg/L) group. The cell migration was analyzed by wound-healing assay, and RT-qPCR was used to measure SQLE mRNA expression. (5) The HepG2 cells with overexpres-sion of SQLE (OE-SQLE) were divided into 5 groups as follows: control group, overexpression negative control (OE-NC) group, OE-SQLE group, OE-NC+pachymaran group, and OE-SQLE+pachymaran group. The mRNA and protein expression levels of SQLE and pyroptosis-related factors were determined by RT-qPCR and Western blot. Colorimetric method and ELISA were used to measure lactate dehydrogenase (LDH), interleukin-1β (IL-1β) and IL-18 levels. The necrosis of HepG2 cells was analyzed by flow cytometry. RESULTS: The SQLE gene was screened through bioinformatics analysis, and its mRNA expression was significantly increased in tumor tissues from HCC patients (P<0. 01). In cell experiments, treatment with 800 mg/L pachymaran for 48 h had a significant inhibitory effect on HepG2 cell viability, and the expression of SQLE mRNA was reduced (P<0. 01). After overexpression of SQLE, the mRNA and protein levels of pyroptosis-related factors, necrotic rate, and LDH, IL-1β and IL-18 levels were significantly decreased (P<0. 05). After treatment with pachymaran, the above indicators were significantly increased (P<0. 05). CONCLUSION: The SQLE is abnormally highly expressed in HCC, and pachymaran can affect the growth of HCC cells by activating the NLRP3/GSDMD pyroptosis pathway through SQLE. [ABSTRACT FROM AUTHOR]

We analyzed the phenomenon which is similar to "node" and " serratula" generated by natural pouring water, restored the phenomenon through a large number of repeated experiment and figure out the causes of the phenomenon with the combination of mechanical analytic modeling and computer simulation. This paper, by using the fluid continuity equation, combined with the appropriate boundary conditions, obtained the exports rate distribution of water in the smooth cylinder, and pointed out that the functional relationship between the node and quantity of flow. We manufactured asymmetric rate distribution to produce rotation of the water column. Finally, through simulation, we improved the credibility of the theory. [ABSTRACT FROM AUTHOR]

This paper is the first attempt to use the non-cooperation game theory to study the strategies of airline carbon tax from EU. Based on inducing four scenarios, non-resistance, refusal of payment, retaliatory duties and creditable threat, Stackelberg game models are developed to find the optimal strategies of countries. The results show that the strategy of refusal of payment, which China is taking step, is superior to the retaliatory duties. The main contribution of this paper is to study a new important international issue, airline carbon tax, summarize four coping strategies and analyze this issue by establishing Stackelberg game models for the first time. [ABSTRACT FROM AUTHOR]

Base on Timoshenko beam and higher-order shear deformation theory, the effects of shear deformation and piezo-electric-elastic coupling are considered. The beam is divided into different regions in order to describe the different sizes and positions of the delamination easily. The governing equations for the flexural wave propagation in the beam are derived based on Timoshenko beam theory. According to the potential distributing simplification for the long wave propagation, the harmonic response of the cantilever beams without the additional potential is gained. The dispersion relations and the closed solution of the displacements and the potentials are obtained. The influence of delamination parameters on displacements and potentials is discussed. [ABSTRACT FROM AUTHOR]

A new type of coal modifying additive (ZBS) was developed to modify the non-coking coal, so it can be used for coking. A series of 40 kg coking-oven experiments were performed and the results show that the quality of the coke deteriorates with the addition of 4% of anthracite and 6% of Long Flame Coal, while it is improved significantly with the ZBS modifying additive. The best improvement is achieved when 0.08% of ZBS is mixed with the non-coking coal, which causes coke properties such as the anti-broken strength of coke (M40) to increase by 5.94%, wear resistance strength (M10) to decrease by 7.12%, coke reactivity index (CRI) to reduce by 4.61% and strength after reaction (CSR) to increase by 11.42%. The quality of the coke obtained is very close to the production index of coking coal and the production requirements can be met. [ABSTRACT FROM AUTHOR]

Microaneurysm is the initial symptom of diabetic retinopathy. Eliminating this lesion can effectively prevent diabetic retinopathy in the early stage. However, due to the complex retinal structure and the different brightness and contrast of fundus image because of different factors such as patients, environment and acquisition equipment, the existing detection algorithms are difficult to achieve the accurate detection and location of the lesion. Therefore, an improved detection algorithm of you only look once (YOLO) v4 with Squeeze-and-Excitation networks (SENet) embedded was proposed. Firstly, an improved and fast fuzzy c-means clustering algorithm was used to optimize the anchor parameters of the target samples to improve the matching degree between the anchors and the feature graphs; Then, the SENet attention module was embedded in the backbone network to enhance the key information of the image and suppress the background information of the image, so as to improve the confidence of microaneurysms; In addition, an spatial pyramid pooling was added to the network neck to enhance the acceptance domain of the output characteristics of the backbone network, so as to help separate important context information; Finally, the model was verified on the Kaggle diabetic retinopathy dataset and compared with other methods. The experimental results showed that compared with other YOLOv4 network models with various structures, the improved YOLOv4 network model could significantly improve the automatic detection results such as F-score which increased by 12.68%; Compared with other network models and methods, the automatic detection accuracy of the improved YOLOv4 network model with SENet embedded was obviously better, and accurate positioning could be realized. Therefore, the proposed YOLOv4 algorithm with SENet embedded has better performance, and can accurately and effectively detect and locate microaneurysms in fundus images.

Objective: To study the expression of hypoxia inducible factor 1α(HIF-1α) of iron-overloaded in irradiated mice and its effect on erythropoiesis., Methods: Twenty mice were randomly divided into 4 groups: Ctrl (control group), IR (irradiation group), IO (irradiation + iron overload group), and RAPA (rapamycin treatment group). The iron overload model was verified. The CFU-E (colony forming unit-erythroid) and BFU-E(burst colony forming unit-erythroid) were cultured; flow cytometry was used to detect the ratios of early stage (Ter119 + CD71 - ) to late stage (Ter119 + CD71 + ) of primitive erythroblasts; RT-PCR was used to detect the mRNA expression of HIF-1α and its related signal molecules in bone marrow cells., Results: The expression of HIF-1α in IR and IO group was significantly higher than that in Ctrl group, and that in IO group was significantly higher than IR group (P<0.05). The ratio of late stage primitive erythroblasts, the number of CFU-E and BFU-E in both IR and IO group were lower than those in Ctrl group, and those in IO group were significantly lower than those in IR group (P<0.05). Compared with Ctrl group, the expression of HIF-1α related signal pathway molecules in both IR and IO group was significantly decreased (P<0.05). Compared with IO group, the expression of HIF-1α and its related signal molecules in RAPA(mTOR inhibitor) group was decreased significantly (P<0.05), the number of BFU-E was increased significantly(P<0.05)., Conclusion: Irradiation induces the increase of HIF-1α and the decrease of the ability of hematopoietic colony formation and the ratio of late stage primitive erythroblasts. Iron overload can aggravate the injury. mTOR inhibitor rapamycin can partially alleviate the injury, suggesting that iron overload can lead to injury of erythropoiesis through HIF-1α.

Based on infrared spectral absorption technique, a carbon monoxide (CO) detection system was developed using the fundamental absorption band at the wavelength of 4.6 μm of CO molecule and adopting pulse-modulated wideband incandescence and dual-channel detector. The detection system consists of pulse-modulated wideband incandescence, open ellipsoid light-collec- tor gas-cell, dual-channel detector, main-control and signal-processing module. By optimizing open ellipsoid light-collector gas- cell, the optical path of the gas absorption reaches 40 cm, and the amplitude of the electrical signal from the detector is 2 to 3 times larger than the original signal. Therefore, by using the ellipsoidal condenser, the signal-to-noise ratio of the system will be to some extent increased to improve performance of the system. With the prepared standard CO gas sample, sensing characteris- tics on CO gas were investigated. Experimental results reveal that, the limit of detection (LOD) is about 10 ppm; the relative er- ror at the LOD point is less than 14%, and that is less than 7. 8% within the low concentration range of 20~180 ppm; the maxi- mum absolute error of 50 min long-term measurement concentration on the 0 ppm gas sample is about 3 ppm, and the standard deviation is as small as 0. 18 ppm. Compared with the CO detection systems utilizing quantum cascaded lasers (QCLs) and dis- tributed feedback lasers (DFBLs), the proposed sensor shows potential applications in CO detection under the circumstances of coal-mine and environmental protection, by virtue of high performance-cost ratio, simple optical-path structure, etc.

Four typical types of green shrubs of Beijing (Euonymus japonicus, Buxus microphylla, Berberis thunbergii cv. atropurpurea, Taxus cuspidate cv. nana) were selected to study their capacities in adsorbing and arresting PM2.5 using both field observations and air chamber simulations. Concurrently, in order to analyze the pollution characteristics of Beijing in winter and spring, the PM2.5 concentrations of December 2012 to May 2013 were collected. Experimental results showed that: From the gas chamber experiments, the ability to adsorb and arrest PM2.5 was in the order of Berberis thunbergii cv. atropurpurea > Buxus microphylla > Taxus cuspidate cv. nana > Euonymus japonicus, mainly due to the differences in leaf characteristics; Outside measurement results showed that the ability to adsorb and arrest PM2.5 was ranked as Buxus microphylla > Berberis thunbergii cv. atropurpurea > Taxus cuspidate cv. nana > Euonymus japonicus. Chamber simulation and outdoor observation showed that Buxus microphylla and Berberis thunbergii cv. atropurpurea had strong ability to adsorb and arrest PM2.5; Meanwhile, the slight differences between the chamber simulation and outdoor observation results might be related to plant structure. Compared to tree species, the planting condition of shrub species was loose, and it greened quickly; By analyzing the Beijing PM2.5 concentration values in winter and spring, it was found that the PM2.5 concentration was particularly high in the winter of Beijing, and evergreen shrubs maintained the ability to adsorb and arrest PM2.5.

Objective: To study the characteristics of EV71 JN200804 strain infection in one-day old BALBI c mjce and to establish a animal model of EV71 infection , and to provide information and technical support for the evaluation of the EV71 vaccine and antiviral medicine., Methods: One-day old BALBic mice were infected with EV71 JN200804 strain through oral( PO) ,intracranial(IC) ,intraperitoneal (IP), intramuscular (IM) routes, respectively. All mice were sacrificed at paralysis of hind limbs and collected organs for viral isolation, RT -PCR and pathological examination, and the electrophysiology were detected before sacrifice., Results: All mice infected through IC, IP and IM routes were paralyzed in hind limbs at 4-5 days and died at 7 days about, the hypokinesia and lethargy of mice were observed through PO routes. The viruses could be isolated and detected in the muscle from mice infected through IC, IP and IM routes and in the spinal cord through IC routes by viral isolation and RT-PCR. The neurogenic and myogenic disorders were detected by electromyography. Histopathologically, the varied pathologic changes were observed in the mouse cerebellum , spinal cord , muscle , heart, lung, liver and kidney., Conclusion: EV71 JN200804 strain can infect one-day old BALBI c mice and induce paralysis of hind limbs, its animal infection model may apply to study of EV71 infection pathogenesis and antiviral medicine, and evaluation of the EV71 vaccine.

Objective: To investigate the neuroprotective effect of mulberry extracts (ME) against Abeta25-35-induced injury in cultured PC12 cells and explore the possible mechanisms involved., Methods: PC12 cells at logarithmic growth phase were divided into the normal control group, Abeta25-35 group (20 micromol/L for 24h) and ME pretreating groups at the concentrations of 25, 50, 100, 200, 400, 800 microg/ml. Cell viability was determined by MTT assay. On that basis, PC12 cells were divided into the normal control group, ME group (200 microg/ml ME incubation alone for 24h), Abeta25-35 group (20 micromol/L for 24h) and ME plus Abeta25-35 group (after pretreatment with 200 microg/ml ME for 24h, cells were exposed to 201 micromol/L Abeta25-35 for another 24h). Intracellular reactive oxidative species (ROS) was measured by using the fluorescent DCFH-DA and the apoptotic cells were observed by Hoechst33342 staining method., Results: (1) The results showed that exposure of PC12 cells to Abeta22-35 (20 micromol/L) for 24h induced notably neuronal injury (P < 0.05) as compared with the control group, while pretreatment with 100 or 200 microg/ml ME almost completely reversed Abeta25-35 induced neuronal injury (P < 0.05). (2) There were no remarkable changes in ROS levels and apoptosis rate of ME group as compared with the control group (P > 0.05). Compared with the control group, exposure to 20 micromol/L Abeta25-35 for 24h resulted in a significant decrease in cell viability (P < 0.05) while increase in ROS levels and cell apoptosis rate (P < 0.05). But pretreatment of PC12 cells with 200 microg/ml ME could counteract ROS formation and inhibit apoptosis (P < 0.05)., Conclusion: These results suggest that mulberry extracts rich in phenolics and anthocyanins could alleviate Abeta25-35-induced injury in PC12 cells, which might be associated with the antioxidative and antiapoptosis effects.

Objective: To observe the difference in the efficacy on the symptoms of bronchial asthma at the chronic persistent stage between acupoint heat-sensitive moxibustion and western medicine with Seretide., Methods: Sixty-four cases were randomly divided into a heat-sensitive moxibustion group (32 cases) and a western medication group (32 cases). In the heat-sensitive moxibustion group, the sensitized points located between Feishu (BL 13) and Geshu (BL 17) or in the region 6-cun lateral from the 1st and the 2nd intercostal spaces of the chest were selected. The heat-sensitive moxibustion was adopted, continuously for 8 days, once per day. In the later 22 days of the 1st month, 12 treatments should be ensured. Two months later, 15 treatments should be guaranteed each month. The time of each treatment was 30 to 90 min. Totally 50 treatments were required. In the western medication group, Seretide inhaler was adopted, one inhalation each time, twice per day, for 3 months totally. The asthmatic symptoms were scored for the patients in two groups and the comparison was made between the two groups., Results: After 3 months of treatment, the asthmatic symptom scores were all improved for the patients in the heat-sensitive moxibustion group and the western medication group as compared with those before treatment (both P < 0.05). In 6 months of follow-up visit, the asthmatic symptom scores in the heat-sensitive moxibustion group were stable, but those in the western medication group were reduced, there was significant difference between the two groups (P < 0.05)., Conclusion: The acupoint heat-sensitive moxibustion effectively relieves the clinical symptoms for the patients with bronchial asthma at the chronic persistent stage. Its efficacy is similar to that of Seretide inhaler. But the long-term efficacy of the heat-sensitive moxibustion is much better.

Objective: To investigate the expression of transient receptor potential vanilloid subtype 1 (TRPV1) in uterosacral ligament and its correlation with pain in endometriosis., Methods: Total of 54 patients undergoing endometriotic lesions excision in uteroscaral ligament by laparoscopy due to pelvic pain were enrolled in this study. According to visual analogue scale (VAS) scores, 27 patients with VAS 5-10 were in group A and 27 patients with VAS 0-4 were in group B. In the mean time, 20 patients with dysmenorrhea without endometriosis (VAS: 0-4) were matched as group C. Specimens (including the sacro-ligaments of 20 women without endometriosis) were immunostained with specific antibodies of TRPV1. Western blot and real time PCR were performed to detect TRPV1 expression in endometriosis lesions and control group., Results: (1) Immunohistochemnistry: the positive area of TRPV1 was found in endometriotic lesions in uterosacral ligament in group A, B and tissue of uterosacral ligament group C. The semi-qualitification of TRPV1 expression were 3 in group A, 1 in group B and 1 in group C by immunohistochemistry staining. There was significantly different expression between group B and group A (P = 0.005) or group C (P = 0.027). (2) mRNA expression:the expression of TRPV1 was 1.84 in group A, 0.80 in group B, 0.24 in group C, respectively. With higher VAS scores, the expression of TRPV1 exhibited increasing trends. The expression of TRPV1 mRNA was higher in group A than that in group B (P = 0.022). There was statistically different expression between group B and group C (P = 0.031). (3) Western blot: the expression of TRPV1 protein was 0.63 in group A, 0.19 in group B, 0.02 in group C. There was significant differences between group A and group B (P = 0.022), and between group B and group C (P < 0.01)., Conclusion: The expression of TRPV1 was correlated with the degree of pain in patient with endometriosis.

Objective: To explore the therapeutic effect of transplantation of bone marrow mesenchymal stem cells (BMSCs) over-expressing Cx43 on heart failure in post-infarction rats., Methods: Sixty SD rats were randomly divided equally into 4 groups: sham group, DMEM/F12 group injected with DMEM/F12, EGFP group with transplanted EGFP transfected BMSCs and Cx43 group with transplanted Cx43 transfected BMSCs. Myocardial infarction model was established by ligating anterior descending branch and the cells were transplanted after 30 minutes. At Week 4 post-infarction, the heart functions of rats were evaluated by echocardiography. After the rats were sacrificed, their tissue samples were collected. The areas of myocardial infarction and the levels of collagen fiber content were measured. And the expressions of EGFR and Cx43 were observed under laser confocal microscopy. The level of Cx43 mRNA was measured by reverse transcription-polymerase chain reaction (RT-PCR)., Results: As compared with the DMEM/F12 group, cardiac function was improved significantly, myocardial infarct area shrunk and collagen fiber content decreased significantly in the EGFP and group in Cx43 groups the. Survival of BMSCs and the formation of gap junction between BMSCs and the host myocardium could be observed under laser confocal microscopy both in EGFP group and Cx43 groups. And the post-infarction, expression of Cx43 mRNA in myocardial tissue decreased significantly in the group DMEM/F12, when compared with sham group (0.18 ± 0.05 vs 0.78 ± 0.14, P < 0.01). There was no significant difference on expression of Cx43 mRNA between DMEM/F12 group and EGFP group (0.18 ± 0.05 vs 0.20 ± 0.09, P > 0.05). The lever of Cx43 mRNA was higher in group Cx43 than in group DMEM/F12 and group EGFP(0.39 ± 0.14 vs 0.18 ± 0.05, P < 0.01; 0.39 ± 0.14 vs 0.20 ± 0.09, P < 0.05)., Conclusion: Transplantation of BMSCs attenuates ventricular remodeling and improves cardiac functions. It may result from the over-expression of Cx43 gene through its effects of improving gap junction remodeling and increasing electro-mechanical coupling between myocardial cells in peri-infarct area.

Objective: To investigate the effects of decompresion through double-incision of foot dorsum on the treatment of osteofascial compartment syndrome of the foot under the monitoring of saturation of blood oxygen., Methods: From January 2000 to June 2007, 26 cases of osteofascial compartment syndrome of the foot were decompressed through double-incision of foot dorsum under the monitoring of saturation of blood oxygen, and relaxation suture or skin grafting were operated within 3 to 10 days after decompressing. Among them, 22 patients were males and 4 were females, with an average age of 36.3 years old ranging from 22 to 68 years. According to AOFAS system, the pain, function, autonomic activities and support were evaluated., Results: All patients were followed-up for from 6 to 43 months with the average of 19 months. All patients were healed. According to AOFAS system, the total scores increased from preoperative (30.4 +/- 8.02) to postoperative (92.5 +/- 5.0) (t = 3.13, P < 0.01); the results were excellent in 21 cases, good in 4 and poor in 1., Conclusion: The patients of fracture-dislocated, swelling and injured in the soft tissue because of severe violence should observed closely on osteofascial compartment syndrome of the foot early. Feet are operated and thoroughly decompressed as soon as it is diagnosed as the compartment syndrome. Osteofascial compartment syndrome of the foot decompressed by foot dorsum double-incision is convenient and satisfied, and the operation is performed by internal fixation if it is displacedly fractured at the same time.

Objective: To investigate the activity of a novel adjuvant consisting of dimethyldioctyldecyl ammonium bromide (DDA) and BCG polysaccharide nucleic acid (BCG-PSN)., Methods: BCG-PSN was extracted by hot-phenol method, and combined with DDA and Mycobacterium tuberculosis fusion antigen AMM (Ag85B-MPT64(190-198)-Mtb8.4) to formulate the Mycobacterium tuberculosis subunit vaccine. Mice were immunized subcutaneously with a 2-week interval between the immunizations (0.2 ml/dose), and humoral and cell-mediated immunity were detected by ELISA and ELISPOT respectively., Results: With the stimulation of Ag85B in vitro, the number of antigen specific IFN-gamma producing spleen lymphocytes were 222 +/- 79, 259 +/- 85, 230 +/- 64 per million respectively in the mice immunized with AMM + DDA + BCG-PSN, AMM + DDA, and BCG. Spleen lymphocytes in these 3 groups produced higher levels of IFN-gamma compared to the groups with the adjuvant of IFA or BCG-PSN alone or without adjuvant upon stimulation with Ag85B (t = 2.923-7.118, P < 0.05). Furthermore, the adjuvant consisting of DDA and BCG-PSN increased the ratio of Ig(2a)/IgG1 than DDA alone (0.125 vs. 0.025). Combined with AMM, the adjuvant DDA and the one consisting of DDA and BCG-PSN induced higher level of immunity than incomplete Freund's adjuvant (IFA), NaCl, and BCG-PSN alone., Conclusion: Mycobacterium tuberculosis subunit vaccine AMM + DDA + BCG-PSN induced a strong Th1-type immune response, and DDA + BCG-PSN, especially DDA promoted the immune response of the M. tuberculosis subunit vaccine in mice.

Objective: To investigate whether intralesional injection of low-dose 5-fluorouracil (5-Fu) and corticosteroid can increase effective rate and decrease recurrence rate of surgically removed auricular keloid., Methods: Eighty-three patients with 166 total ear keloids were studied, including 79 keloids of helix and 87 of earlobe with an average disease history of 2.9 years. Ear piercing accounted for 92.7% for etiology. Keloids were first surgically removed in order to keep the normal auricular shape. Then intralesional injection of low-dose of 5-Fu and corticosteroid was given 3 - 4 weeks after surgery and every 4 weeks afterwards. If no recurrence, the injection was given every 2 to 3 months with gradually tapered dose. The evaluation standard includes: patients with no recurrence 6 months after stopping drug injection and maintaining of the normal auricular shape are considered as "cure"; those remain having minor auricular deformity 6 months post-surgery with no recurrence and needing further treatment are considered as "effective"., Results: The treatment time period for 83 patients was 2 - 26 months (mean: 7 months). The patients were followed up for 0 - 49 months (mean: 9 months) after withdrawing drug injection. There were 39 cured cases (47.0%) and 44 effective cases (53%). The total effective rate was 100%. Normal auricular shapes were achieved in 77 cases (92.7%) ., Conclusion: Surgical removal of auricular keloid followed by intralesional injection of low-dose 5-Fu and steroid is an effective method to treat auricular keloid and prevent its relapse.

Background: CAlthough there are increasing reports on localization of tracheal stem cells, the mechanism of proliferation and differentiation of tracheal stem cells remains unclear., Methods: In this study, we developed an ex vivo model of murine tracheal epithelial injury and regeneration induced by 5-FU. The regeneration process of murine tracheal epithelium was observed and analyzed by morphological, immunofluorescence and microarray methods., Results: After treatment with 5-FU, the differentiated mature cells were dead and desquamated. Only a few cells remained in G0 phase and located on the basement membrane. After being put back in normal culture media, the cells became flat, cubic and restored as pseudostratified epithelium at last. These G0 phase cells were ABCG2 positive. We detected the differences of stem cell genes between normal tracheal epithelial cells and regenerated epithelial cells at 24 h and 48 h after injured by 5-FU using stem cell differentiation gene microarray. At 24 h, 8 genes were up-regulated and 31 genes were down-regulated. At 48 h, 5 genes were up-regulated and 42 genes were down-regulated., Conclusions: The differently expressed genes were mainly associated with cell cycle regulation, intercellular junction, FGF, BMP, Notch and Wnt signaling pathways, which suggested these alterations might be closely associated with the proliferation and differentiation of tracheal stem cells.

Objective: To explore the feasibility of electroporation mediated gene transfer in rat incisional wound., Methods: 12 Sprague-Dawley rat's dorsal skins were electroporated (800 voltages in amplitude with 6 square wave pulses, each lasting 20 milliseconds with 200 millisecond interval) after injection of plasmid DNA (1 microg/microl, in 100 microl PBS) containing enhanced green fluorescence protein (EGFP) gene. Electroporated skins were incisionally wounded 24 hours after electroporation. Specimens were harvested at day 2, 4, 6, 14, then EGFP expression in dermis was observed and quantitatively analyzed with integrated optical density (IOD) followed by H&E staining., Results: Electroporation can mediate EGFP expression in epidermis, dermis and panniculus muscle. The expression level in dermis was the highest at day 2 (IOD = 3.50 +/- 1.45) and disappeared at day 14. EGFP expression was not found in dermis if no electroporation applied after plasmid injection (IOD = 0)., Conclusion: Electroporation can mediate plasmid gene expression in incisional wound efficiently and widely.

The genetic diversity of 279 indivdiuals from 10 populations in Shandong Province was investigated using inter-simple sequence repeat (ISSR) markers. As a result, 116 bands were amplified by 10 informative and reliable primers, of which 101 were polymorphic loci. A relatively high level of genetic diversity was revealed: PPL = 87.07, He = 0.2697, H0 = 0.3999 (at the species level); PPL = 64.58, He = 0.2004, H0 = 0.3010 (at the population level). A higher level of genetic differentiation was detected among populations with Nei's G(ST) analysis and the analysis of molecular variance (AMOVA; G(ST) = 0.2414, F(ST) = 0.2224). Habitat fragmentation and gene flow may result in genetic differentiation. UPGMA cluster analysis indicated that the four populations from Linshu, Junan, Tancheng and Feixian grouped together, whereas Laiyang populations clustered in an isolated clade. The results showed that a mixed mating system was possibly the main factor influencing the genetic structure of this species. These results, combined with other information about Castanea mollissima, may provide a valuable basis for proposing conservation strategies.