Objective To investigate the effect of silk fibroin scaffolds seeded with adipose mesenchymal stem cells (ADMSCs) in remodeling fiber for tunica albuginea defect in rabbits.Methods Fifty-six New Zealand rabbits were divided into 4 groups according to the random number table after a defect was created in the tunica albuginea:Group A (the defect was repaired with silk fibroin),Group B (with silk fibroin seeded with ADMSCs),Group C (with autologous tunica vaginalis) and Group D (left unrepaired),with 14 rabbits per group.Tunica albuginea sections were obtained for HE staining,Sirius red staining,Hart staining and immunofluorescence staining of macrophages at 6 and 12 weeks after surgery.Results At 12 weeks after surgery,HE staining revealed chaotically distributed new fiber ingrowth in Group D and orderly ingrowth in Groups A,B,and C.At 12 weeks after surgery,Sirius red staining revealed mean area of type Ⅰ collagen fibers was greater than that of type Ⅲ in Groups A (98 780 ±4 190 vs 51 177 ±5 464),B(94 855 ±9 010 vs 50 815 ±3 895),and C(99 860 ±6 015 vs 50 948 ± 6 595),but the difference in area of collagen fiber of the same type was insignificant among the three Groups.Moreover,less type Ⅰ collagen fibers (79 386 ±2 237) and more type Ⅲ collagen fibers (85 278 ± 2 645) were observed in Group D compared with other three Groups (P ＜ 0.01).At 12 weeks after surgery,Hart staining showed the mean area of elastic fibers in Groups A,B,C,and D was 2 805 ± 90,2 849 ±84,3 068 ±485,and 1 961 ±96 respectively.There was no statistical difference between Groups B and C,but less amount of fibers was observed in Group A (P ＜ 0.01) and least amount was observed in Group D (P ＜0.01).At 6 weeks after surgery,the number of infiltrating macrophages in Groups A,B,C and D was 4.10 ± 0.87,3.80 ± 0.78,3.70 ± 0.94,and 6.80 ± 1.63 respectively.There was no statistical difference in the number of macrophage infiltration among Groups A,B and C,but all were lower than that in Group D (P ＜ 0.01).Conclusion Silk fibroin seeded with ADMSCs is comparable to autologous grafts for repair of tunica albuginea defect in rabbits.

OBJECTIVETo investigate the effects of ulinastatin on lung injury in hemorrhagic shock rats.

METHODSTwenty-four normal SD rats were randomly divided into 3 groups (n=8), namely the control group, hemorrhagic shock group (group H) and ulinastatin group (group U). In group H and group U, blood was drawn from the femoral artery over a period of 10 min until a mean arterial pressure of 40 mmHg was obtained. Controlled hypotension was then maintained at 40-/+5 mmHg for 60 min by blood drawing or infusion when necessary. All the blood drawn and an equivalent volume of Ringer lactate solution were subsequently infused for resuscitation. Four hours after the resuscitation, the activity of superoxidedismutase (SOD), content of malondialdehyde (MDA), expression of heme oxygenase-1 (HO-1), wet to dry weight ratio (W/D), and pathologic changes of the lung tissues were measured or observed.

RESULTSCompared with those in the control group, the content of MDA, expression of HO-1 and W/D increased significantly in both group H and group U (P<0.05); these indexes in group U were significantly lower than those in group H (P<0.05). The activity of SOD in group U was significantly lower than that in the control group (P<0.05) but higher than that in group H (P<0.05). Optical microscopy demonstrated milder inflammatory cell infiltration and interstitial edema in the lung tissues in group U than in group H.

CONCLUSIONUlinastatin can lower the content of MDA, W/D and the expression of HO-1, increase the activity of SOD, and reduce histological lung injury in rats with hemorrhagic shock.

Animals ; Glycoproteins ; pharmacology ; Heme Oxygenase-1 ; metabolism ; Lung Injury ; etiology ; prevention & control ; Male ; Malondialdehyde ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic ; complications ; metabolism ; Superoxide Dismutase ; metabolism

To identify the active constituents in vitro and blood-absorbed ingredients in vivo from Yin Chen Hao decoction provides scientific evidence for probing its prevention and treatment mechanism on acute liver injury. An ultrahigh performance liquid chromatography quadrupole-time of flight-mass spectrometry (UPLC-QTOF/MS) method was applied for analysis of Yin Chen Hao decoction and the serum samples of mice with con-A induced acute liver injury after preventive oral administration for 14 days (the use of all laboratory animals in this study was approved by the Ethics Committee of the Naval Medical University, 19YF1459400). A total of 90 chemical constituents were identified from Yin Chen Hao decoction, mainly were flavonoids, terpenoids, tannins, quinones. 5 prototype compounds were identified in the serum, including chrysophanol, deoxyrhapontin-8-O-gallate, mussaenosidic acid, herniarin, emodin. The established UPLC-QTOF/MS method could efficiently and sensitively identify the constituents in vitro and blood-absorbed ingredients of Yin Chen Hao decoction, primarily clarify the material basis of its hepatoprotective effect, and provided a scientific basis for the quality marker selection and the pharmacodynamic material basis research on the decoction.

Traditional Chinese medicine(TCM) and its components play a role in the field of anti-hepatocarcinoma. The definition of its mechanism of action in autophagy contributes to the development of TCM in the field of anti-hepatocarcinoma. In this paper, we summarized reports on the autophagy of liver cancer cells induced by TCM and its active ingredients, including those on promoting apoptosis and cycle inhibition induced by autophagy and inhibiting autophagy to block tumor cell cycle, but with a lack of systematic summarization. In this paper, according to different effect of TCM on autophagy induced by hepatocarcinoma, the TCM and its components were inductively analyzed in four aspects:inducing killing autophagy, inhibiting protective autophagy, inducing protective autophagy in liver cancer, and inducing unclear autophagy. According to the findings, TCMs and components that cause killing autophagy can inhibit the occurrence of autophagy, arrest cell cycle, induce cell senescence or promote apoptosis. TCMs and components that inhibit protective autophagy can inhibit protective autophagy and hepatoma cell proliferation. TCMs and components that induce protective autophagy have a significant anti-hepatocarcinoma effect, shall be considered to be combined with autophagy inhibitors to enhance the lethality of drugs on liver cancer cells, and become a new way for such drugs to treat liver cancer. TCMs and components with an unclear inductive effect shall be first identified for their type of autophagy, then combined with autophagy agonists or blockers according to the type of autophagy to enhance their anti-liver cancer effect, and provide a new clinical therapeutic approach for liver cancer. In the aspect of autophagy, this study not only reveals the molecular mechanism of anti-hepatocarcinoma of TCM, but also makes it a new way to study anti-hepatocarcinoma by TCM.

Objective To construct an adenoviral vector containing mouse Hes1 gene, observe its expression in the hippocampus of adult mice and build a basis for further investigation of Hes1 gene in adult neurogenesis. Methods The restriction endonuclease was used to digest plasmid pEGFP-mHes1 and pDC316, and then, the products were recovered and connected by T4 DNA ligase and the shuttle plasmid pDC316-mHes1 was constructed which was identified by the method of PCR and EcoRI+HindⅢ digestion. After that, the shuttle plasmid pDC316-mHes1 was cotransfected into 293 cells with the adenovirus skeleton plasmid pBHGlox_E1,3Cre to obtain the produced replication defective recombinant adenovirus Ad5-mHes1. Then, the recombinant adenovirus could be further amplified and purified. The report recombinant adenoviruses were Ad5-EGFP containing enhanced green fluorescent protein (EGFP).Then, Ad5-mHes1 and Ad5-EGFP were stereotactic injected into the hippocampus of the adult C57BL/6 mice and their expressions in the hippocampus were detected. Western blotting was used to detect the Hes1 protein level 7 d after the injection. Fluorescent microscopy was used to observe the expression of EGFP in the hippocampus. Results The experimental results of identification by the methods of PCR and EcoRI+HindⅢ digestion were in accordance with the anticipated results, and the sequences were also the same with mHeslCDS sequences; Hes1 gene was expressed in the hippocampus of both the PBS injection group and the Ad5-mHes1 injection group 7 d after the injection, and the expression of Hes1/GAPDH in Ad5-mHes1 injection hippocampus (0.705 ±0.128) was statistically different as compared with that in PBS injection group (0.363±0.053, P＜0.05). Ad5-EGFP strongly expressed in the granular cell layer and subgranular zone (SGZ) of dentate gyrus. Conclusion The adenoviral vector of mouse Hes1 gene is successfully established and Hes1 gene is expressed in the hippocampus of C57BL/6 adult mice.

OBJECTIVETo investigate levels of antibodies against type A and type C influenza viruses and those against the 2009 H1N1 influenza A virus (before and after the 2009 H1N1 pandemic) among residents in Wuxi. To compare levels of antibodies against the 2009 H1N1 influenza virus (one year after the pandemic) in the unvaccinated population with those in the population who received vaccine.

METHODSSerum samples were collected from subjects (aged 1-60 years) during September 2008 to May 2009, and during September 2010 to January 2011. Also collected were serum samples from adults who had received vaccines for pandemic (H1N1) 2009 for one year. Antibody response to influenza viruses was measured using hemagglutination inhibition (HI) assay. Seropositivity rate, seroprotection rate and geometric mean titer (GMT) were compared for each age group during different periods.

RESULTSBefore the outbreak of the 2009 H1N1 pandemic, seropositivity rate, seroprotection rate and GMT among the study subjects in were 2.86% (4/140), 0.71% (1/140) and 5.23, respectively. One year after the outbreak, seropositivity rate, seroprotection rate and GMT among the study subjects were 66.33%, 37.76% and 19.17, respectively. Among them, adult subjects showed 50.00% seropositivity rate, 19.44% seroprotection rate and 13.09 GMT, while adult subjects who had received vaccine for one year showed 61.36% seropositivity rate, 22.73% seroprotection rate and 14.14 GMT. No significant difference was observed between these two populations (P > 0.05 for all three indexes). Furthermore, before the outbreak of the 2009 H1N1 pandemic, levels of antibodies against seasonal influenza viruses among the study subjects were as follows: for H1N1 virus, seropositivity rate, seroprotection rate and GMT were 55.00%, 35.00% and 16.90, respectively; for H3N2 virus, seropositivity rate, seroprotection rate and GMT were 86.40%, 84.30% and 58.56, respectively.

CONCLUSIONOne year after the 2009 H1N1 influenza A virus had spread to Wuxi, the population levels of antibodies against this virus have approached those against seasonal influenza viruses, as reflected by seropositivity rates, seroproection rates and GMT. Moreover, considerable levels of antibodies against seasonal influenza viruses were observed in populations, indicating no seasonal influenza outbreak would occur recently.

Adolescent ; Adult ; Antibodies, Viral ; blood ; immunology ; Child ; Child, Preschool ; China ; Female ; Hemagglutination Inhibition Tests ; Humans ; Infant ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza Vaccines ; immunology ; Influenza, Human ; blood ; immunology ; prevention & control ; virology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo observe the preventive and therapeutic effect of advanced airway management on pulmonary infection in patients with inhalation injury after tracheotomy.

METHODSfourteen burn patients with inhalation injury admitted to our hospital from January 2001 to December 2004 were enrolled as control (C) group, and they were treated with conventional systemic therapy and management of airway. Twenty-seven burn patients with inhalation injury admitted to our hospital from January 2005 to October 2009 were enrolled as advanced (A) group, and they were treated with conventional systemic therapy and advanced airway management, including bedside isolation of airway, fixation of both oxygen supply tube and humidifying tube, humidification in specific body position, thinning of sputum, lavement of airway and procedural sputum elimination, steam inhalation combined with medicine, and suction of sputum with interrupted negative pressure. Result of bacterial culture of sputum (the 7th day after tracheotomy) and chest X-ray (at admission and the 7th day after tracheotomy), pulmonary infection, change in blood gas analysis index and oxygen saturation (SO(2)), (within 7 days after tracheotomy), and the number of patients curd in 2 groups were observed and compared.

RESULTS(1) Positive result of bacterial culture of sputum was observed in 11 (78.6%) patients in C group and 12 (44.4%) patients in A group. The difference between them was statistically significant (chi(2) = 4.36, P < 0.05). The main bacterium detected was Pseudomonas aeruginosa. (2) Pneumonia was suspected in 7 patients (25.9%) in A group by chest X-ray, which was obviously fewer than that in C group (8 Cases, 57.1%, chi(2) = 3.87, P < 0.05). The result was in accordance with the diagnosis of pulmonary infection. (3) No CO(2) retention, SO(2) and PaCO(2) abnormality caused by asphyxia was observed in 2 groups, PaCO(2) value in A group was close to that in C group (t = 0.89, P > 0.05). (4) In C group, 9 (64.3%) patients were cured, 5 patients died of pneumonia, wound sepsis, and MODS. In A group, 25 (92.6%) patients were cured, 2 patients died of MODS. Number of cure was obviously larger in A group than in C group (chi(2)= 5.22, P < 0.05).

CONCLUSIONSThe advanced airway management has better effects on isolation and humidification of airway, and thinning, drainage, and elimination of sputum. And it can decrease the probability of blind suction and injury to airway, and it prevents pulmonary infection following tracheotomy.

Adult ; Airway Management ; Burns, Inhalation ; therapy ; Female ; Humans ; Lung Diseases ; etiology ; prevention & control ; Male ; Middle Aged ; Respiratory Tract Infections ; etiology ; prevention & control ; Tracheotomy ; Young Adult

OBJECTIVETo explore the relationship between polymorphisms of FAS and FASL genes and genetic susceptibility of silicosis.

METHODSA case-control study was conducted. The case group was 183 male patients with silicosis and the control group was 111 male silica-exposed but without silicosis miners. Data on total dust concentrations was collected to estimate cumulative total dust exposure (CTE) of each subject and each person's characteristics and work history were obtained from questionnaire. Polymerase chain reaction re-strained fragment length polymorphism technique (PCR-RFLP) was applied to detect the single nucleotide polymorphisms (SNPs) of FAS-1377, FAS-670 and FASL-844. Associations between polymorphisms and risk of silicosis and stages, interactions between polymorphisms, between polymorphisms and CTE and smoking and haplotypes were analyzed.

RESULTSThere were no differences in the FAS-1377, FAS-670 and FASL-844 genotypes between the case group and the control group (P > 0.05). No association was observed between FAS-1377, FAS-670 and FASL-844 polymorphisms and silicosis and stages (P > 0.05). The frequencies of FAS-1377G/-670G haplotype in the cases (9.6%) were higher than those in the controls (3.6%) (P < 0.05). No interactions between the polymorphisms of different genes, the gene polymorphism and the total accumulative total dust, the gene polymorphism and smoking were observed (P > 0.05).

CONCLUSIONFAS-1377, FAS-670 and FASL-844 polymorphisms are not susceptible factors of silicosis. The FAS-1377G/-670G haplotype might be a susceptibility marker of silicosis.

Aged ; Aged, 80 and over ; Case-Control Studies ; Fas Ligand Protein ; genetics ; Genetic Predisposition to Disease ; Humans ; Male ; Middle Aged ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Silicosis ; genetics ; fas Receptor ; genetics

OBJECTIVETo deliver the naked genes into cells through the bioeffects of cell membrane porous produced by low-frequency ultrasound (US) and to investigate the safety by determining the threshold of cell damage and membrane permeability.

METHODSThe suspension of red cells from chickens, rabbits, rats, and S180 cells was exposed to calibrated US field with different parameters in still and flowing state. Laser scanning confocal microscopy, fluorescent microscopy, scanning electron microscopy, flow cytometry and spectrophotometry were used to examine cell morphology, membrane permeability, enzymes, free radicals, naked gene expression efficiency, threshold of cell damage and cell viability.

RESULTSThe plasmid of green fluorescent protein (GFP) as a reporter gene was delivered into S180 cells under optimal conditions without cell damage and cytotoxicity. The transfection rate was (35.83 +/- 2.53)% (n = 6) in viable cells, and the cell viability was (90.17 +/- 1.47)% (n = 6). Also, malondialdehyde, hydroxyl free radical, alkaline phosphatase, and acid phosphatase showed a S-shaped growth model (r = 0.98 +/- 0.01) in response to the permeability change and alteration of cell morphology. The constant E of energy accumulation in US delivery at 90% cell viability was an optimal control factor, and at 80% cell viability was the damage threshold.

CONCLUSIONUS under optimal conditions is a versatile gene therapy tool. The intensity of GFP expression in US group has a higher fluorescent peak than that in AVV-GFP group and control group (P < 0.001). The optimal gene uptakes, expression of gene and safety depend on E, which can be applied to control gene delivery efficiency in combination with other parameters. The results are helpful for development of a novel clinical naked gene therapeutic system and non-hyperthermia cancer therapeutic system.

Animals ; Cell Membrane ; chemistry ; metabolism ; ultrastructure ; Cell Survival ; genetics ; Cells, Cultured ; Chickens ; DNA Damage ; genetics ; Free Radicals ; metabolism ; Genes, Reporter ; genetics ; Green Fluorescent Proteins ; genetics ; Malondialdehyde ; metabolism ; Permeability ; Plasmids ; genetics ; Porosity ; Rabbits ; Rats ; Risk Assessment ; Transfection ; methods ; Ultrasonics

miR-34a is a conserved microRNA highly expressed in the brain. It is thought to play critical roles in regulating many aspects of brain development and function, such as neural stem cell proliferation and differentiation, neuronal migration and apoptosis, fear memory consolidation, etc. However, the assessment of its function was mainly conducted through vector-mediated overexpression and miRNA sponge or antagomir-mediated functional suppression, therefore may suffer from nonspecific off-target effects or incomplete inactivation. We thus analyzed mouse model with a targeted deletion of miR-34a which completely abolishes its expression. To our surprise, loss of miR-34a led to neither an obvious change in brain size, morphology or cortical lamination, nor impaired marker gene expression in major excitatory and inhibitory neuron types in the neocortex. In addition, miR-34a ablation did not affect fear memory formation or consolidation, as well as the anxiety or depression related behavior. However, the performance of mice in rotarod assay was significantly affected, suggesting a defect in motor activity in miR-34a deficient mice. As neocortical parvalbumin (PV) neurons are known for high level miR-34a expression, we also tested the effect of PV-Cre-mediated conditional miR-34a deletion. Similar as germline deletion, PV neuron specific miR-34a deletion did not affect cortical lamination or PV expression in the neocortex. Our studies suggest that, although miR-34a may be involved in regulating certain aspects of brain development or function, such as motor activity, it does not play a significant role in regulating brain morphogenesis, cortical lamination or neocortical neuron subtype specification, and it is also dispensable for fear memory formation, expression and consolidation.