Objective To construct recombinant lentivirus silence vector aiming at rictor gene in mTORC2 specific protein, and to investigate its regulation on mTORC2/SGK1 signal pathway and the effect on pulmonary alveolar epithelial sodium ion chan-nel,as well as the role in acute respiratory distress syndrome(ARDS)and acute lung injury.Methods The interfering vector plas-mid and empty vector plasmid of target gene rictor were constructed,which and the lentivirus packaging system were co-transfected to 293T cells.The viral supernatant was collected,centrifuged,concentrated and purified for obtaining recombinant lentivirus.The virus titer was detected and the virus was infected to A549 cells.Stable cell lines were screened.RT-PCR was used to confirm the silencing situation of target gene rictor.The expression situation of various signal indexes in this pathway was detected by PCR and Western blot.Results The recombinant lentivirus of silence gene rictor was successfully constructed and transfected to A549 cell for obtaining stable cell lines.Compared with blank and control groups,the mRNA levels of rictor,downstream SGK1 andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased (P <0.05 ).Meanwhile,the protein levels of rictor,downstream SGK1,P-SGK andα-,β-andγ-ENaC in the shRNA-rictor group were significantly decreased compared with the other two groups(P<0.05).Conclusion Silence rictor gene has the obvious regulation effect on mTORC2/SGK1 signal pathway,meanwhile affects the expression of pulmonary alveolar epithelial cellular α-,β-and γ-ENaC at gene and protein level.It is speculated that mTORC2/SGK1 may be an important signal pathway for regulating the clearance capacity of pulmonary alveolar epithelial cells on pulmonary alveolar fluid and simultaneously affecting the pulmonary edema formation.

0.05). But there was more BMP in the treated group of periodontitis than the other groups(P

Objective: To demonstrate the effect of structured and autonomous sports activities on children s basic motor skills in a preset outdoor field for a certain time and frequency, and to provide a reference for improving children s development of basic motor skills. Methods: Selected 141 typical developing children from 4 public kindergartens in Nanning, and randomly divided them into experimental group(71) and control group(70). The experimental group participated in physical activities for 10 weeks (3 times a week, 1 hour each time) in the preset outdoor field (each physical activity was composed of 30 minutes of autonomous physical activity and 30 minutes of structured physical activity), while the control group had no intervention of site preset and specially designed activities. Before and after the experiment, 9 basic motor skills (including 3 fine motor skills and 6 gross motor skills) were tested. Results: There were significant differences in double hand throwing, single leg balance (left), single leg balance (right), swing bridge balance and beam walking before and after intervention in experimental group(t=7.82,4.25,9.69,-10.01,-28.28,P<0.05), but there was no statistical significance in the improvement of heel toe coordinated walking(t=-0.60,P>0.05). There was no significant improvement in coin throwing (right hand), coin throwing (left hand) and square stacking(t=0.91,1.03,0.42,P>0.05), while in the control group, except for the balance of swinging bridge(t=4.27,P<0.05) and heel and toe walking(t=3.96,P<0.05), the improvement of other basic motor skills was not statistically significant(t=1.21, 0.74, 0.31, 0.97,P>0.05). Conclusion The quality of sports environment has a potential impact on the basic motor skills of preschool children. The basic motor skills of the pre-school children can be greatly improved by conducting structural and autonomous physical activities with a certain time and frequency.

OBJECTIVE: The study was carried out on time limit of ACP and sperm positive-detected in mixed spot. METHODS: 600 vaginal swabs from living body after sexual intercourse were tested by routine and three-blindness methods. RESULTS: The longest time limit of ACP which was positively detected is 255 h, average time limit is 52 h, 41.3% of those samples was ACP-detected in positive degree (++). The longest time limit of sperm positively detected is 132 h, average time limit is 29 h. The number of sperms observed in 34.3% of those samples with positive result, is from 6 to 10/HP. CONCLUSION This study is helpful to forensic identification of sex cases.
Acid Phosphatase/analysis* ; Coitus ; Female ; Forensic Medicine ; Humans ; Male ; Spermatozoa ; Time Factors ; Vaginal Smears

BACKGROUNDThe Taq/B, Msp/ and I405V polymorphisms of cholesteryl ester transfer protein (CETP), an important regulatory factor of lipid metabolism, have been attracted much more attention by the researchers. In this study, we investigated the associations between these 3 polymorphisms of CETP gene and variations in plasma lipid and lipoprotein levels in patients with coronary heart disease (CHD).

METHODSGenomic DNA was extracted from leukocytes of 203 CHD patients and 100 control subjects using the salting out method. Genotyping of the CETP gene was performed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques. Statistical analysis was conducted using the SPSS 10.0 software package.

RESULTSThe distribution of allele and genotype frequencies of the Taq/B, MspI, and I405V polymorphisms was similar in the CHD patient group and the control group. The B1B1 genotype of the Taq/B polymorphism was associated with significantly higher TC (P=0.039) and LDL-C (P=0.044) levels than the B2B2 genotype in CHD patients, and with significantly higher LDL-C (P=0.034) levels than the B2B2 genotype in controls. Homozygotes of the I405V polymorphism exhibited significantly higher HDL-C levels than VV homozygotes among control subjects (P=0.023). In male CHD patients with unambiguously assigned haplotypes, B2-M2-V/B2-M2-I patients demonstrated significantly higher HDL-C concentrations than B1-M2-V/B1-M2-I (P=0.023) and B1-M2-V/B1-M2-V patients (P=0.047).

CONCLUSIONSGenetic variations in the CETP gene may account for a significant proportion of the differences in plasma lipid and lipoprotein concentrations among the general population. The B1B1 genotype of the Taq/B polymorphism is probably a genetic risk factor for CHD in the study population.

Adult ; Aged ; Carrier Proteins ; genetics ; Cholesterol Ester Transfer Proteins ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Coronary Disease ; blood ; genetics ; Female ; Gene Frequency ; Glycoproteins ; genetics ; Humans ; Lipids ; blood ; Male ; Middle Aged ; Polymorphism, Genetic

Objective To summarize the clinical features of vascular malformations complicated with airway obstruction and to evaluate the therapeutic methods of these disease. Methods Forty-seven children with airway obstruction and dyspnea (25 males, 22 females) were treated from Jun 1985 to Dec 2007, and their clinical data were retrospectively analyzed. Among 47 patients, there were 27 cases of venous malformations,17 cases of macrocystic lymphatic malformations, and 3 cases of microcystic lymphatic malformations. Injection with absolute alcohol were performed in 20 patients with venous malformations, whereas both sugery and injection were performed in 7 patients with extensive or multiple lesions. Seventeen patients with macrocystic lymphatic malformations were treated with pingyangmycin injection. While surgery combined with pingyangmycin injection were used in other 3 patients with microcystic lymphatic malformations. According to the degree of airway obstruction and therapeutic conditions, tracheal intubation was performed in 27 patients, urgent preoperative tracheotomy was performed in 3 patients, prophylactic tracheotomy was performed in 2 patients, and postoperative tracheotomy was performed in 1 patient. Results Tracheal intubation was remained for 24 to 48 hours in 30 patients, whose intubation was removed successfully in 29 patients except 1 patient who occurred dyspnea after removal of tracheal intubation resulting in tracheotomy. Tracheal cannula was successfully removed in all 6 patients 3 weeks to 4 months after the tracheotomy. There were 9 patients treated once, whereas injections were repeated 2 to 5 times in 38 patients. Necrosis of mucosa occurred in 2 cases after the injection with absolute alcohol, while temporary hemoglobinuriaone occurred in 1. There were 5 cases of light or mediate fever after the pingyangmycin injection who recovered well after the symptomatic treatment. Follow-up lasted 1 to 23 years, 38 patients cured, 9 patients valid, and no patient invalid. Conclusions It is suggested that sclerotherapy should be the first choice in the treatment of vascular malformations complicated with airway obstruction, in which absolute alcohol should be used in venous malformations compared to pingyangmycin in lymphatic malformations. Combined therapy should be carried out in patients with extensive lesions in order to shorten the course of treatment and to get good therapeutic result.

Objective To evaluate the short-term results and safety of propranolol for the treatment of infantile parotid hemangioma.Methods Oral propranolol was administered to 17 infants with parotid heamngiom at a dose of 1.0-1.5 mg per kilogram of body weight per day.The patients were revisited once a week.The changes of the tumor size, texture and colour were monitored and recorded at a regular interval.The adverse effects after medication were observed and managed accordingly.The short-term results were evaluated using a 4 scales system.Results Among the 17 patients treated, the follow-up time was 5 to 10 months.The overall response was scale I in 0 patient, scale Ⅱ in 0 patients, scale Ⅲ in 5 patients, and scale Ⅳ in 12 patients.No serious adverse effects were encountered.Conclusions Oral propranolol at a lower dose is a safe and effective method for the treatment of infantile parotid heamngioma.The short-term results were excellent and the side effects minimal.

Background Infantile hemangioma (IH) is the most common benign tumor in children with prevalence in the face and neck.Various treatment options including oral propranolol have been described for IH,but the mechanism of drugs remains enigmatic.The aim of this study was to investigate the pathogenesis and establish a reliable in vivo model of IH which can provide platform for drug exploration.Methods Stem cells from the proliferating hemangiomas (HemSCs) were isolated by CD133-tagged immunomagnetic beads.Their phenotype and angiogenic property were investigated by flow cytometry,culturing on Matrigel,real-time polymerase chain reaction (PCR),immunofluorescent staining and injection into BALB/c-nu mice.Results HemSCs had robust ability of proliferating and cloning.The time of cells doubling in proliferative phase was 16 hours.Flow cytometry showed that HemSCs expressed mesenchymal markers CD29,CD44,but not endothelial/hematopoietic marker of CD34 and hematopoietic marker CD45.The expression of CD105 was much lower than that of the reported hemangioma derived or normal mesenchymal stem cell (MSC).Real-time PCR showed that the mRNA levels of vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF) and matrix metalloproteinase-1 (MMP-1) of HemSCs were higher than that of neonatal human dermal fibroblasts (NHDFs) and human umbilical vein endothelial cells (HUVECs).After HemSCs were cultured on Matrigel in vitro,they formed tube-like structure in a short time (16 hours) and differentiated into endothelial cells in 7 days.After 1-2 weeks of implantation into immunodeficient mice,HemSCs generated glucose transporter 1 positive blood vessels.When co-injected with HUVECs,the vascularization of HemSCs was greatly enhanced.However,the single implantation of HUVECs hardly formed blood vessels in BALB/c-nu mice (P ＜0.05).Conclusions HemSCs may be some kinds of primitive mesoderm derived stem cells with powerful angiogenic ability,which can recapitulate human hemangioma by co-injecting into immunodeficient mice with HUVECs.

BmNPV bacmid constructed recently and Red recombinant system were used to rapidly disrupted Bombyx monri nucleopolyhedrovirus (BmNPV) orf60 in Escherichia coli (E. coli) BW25113. BmNPV bacmid isolated from E. coli BmDH10Bac was electroporated into E. coli BW25113, which harbors plasmid pKD46 encoding lamda Red recombinase,to produce E. coli BW25113-Bac, which could be used for gene deletion of BmNPV. A linear fragment was amplified by PCR from plasmid pKD3 (containing a chloramphenicol acetyltransferase gene cat) using a pair of primers with length of 63bp,which had 45 bp homologous to the orf60 gene and 18bp homologous to cat sequences. The linear fragment was electroporated into E. coli BW25113-Bac and homologous recombination occurred between the linear fragment and orf60 with the help of lamda Red recombinase. Three specific primer pairs were used to confirm the replacement of orf60 by cat gene. Western blot analysis showed that orf60 was not expressed in BmN cells infected with knockout bacmid.
Animals ; Bacteriophage lambda ; enzymology ; genetics ; Bombyx ; virology ; Electroporation ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Gene Knockout Techniques ; Genes, Viral ; genetics ; Nucleopolyhedrovirus ; enzymology ; genetics ; Open Reading Frames ; genetics ; physiology ; Recombinases ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

Ultrasound therapy is a new type of therapy technology, there are hyperthermia and high intensity focused ultrasound surgery (HIFUS). Compared with a single frequency system, mutiple frequency system has an additional function to combine power patterns of different frequencies. This function increases the availability of power patterns to treat tumors of various shapes and depths. Therefore, we have in this article proposed a system with the ability to drive ultrasonic phased arrays of multiple resonant frequencies for ultrasound hyperthermia and HIFUS. The results show that this system is able to (1) drive multi-element applicators or phased arrays of a single resonant frequency through the multichannel power amplifiers, (2) concurrently drive transducer with different resonant frequencies, (3) adjust the relative phase and output power of each channel for the scanning ultrasonic focus, and (4) operate each channel with good output stability.
Equipment Design ; Humans ; Neoplasms ; therapy ; Transducers ; Ultrasonic Therapy ; instrumentation