Adolescent ; Aminolevulinic Acid ; pharmacology ; Female ; Humans ; Porphyria, Acute Intermittent ; physiopathology

OBJECTIVE: In order to understand which kind of function genes play an important role for estimating wound age, the variation of difference genes' mRNA expression were compared after injury. METHODS: The mRNA expression levels of seven candidate genes (ICAM-1, NF-κB, MX2, MT1, MT2, sTnI, and Cox6c) were analyzed in contused rat skeletal muscle at different time points using real-time fluorescent quantitative PCR (RT-qPCR). The raw Ct values were normalized relative to that of RPL32 mRNA, and converted to standard Ct values. At each time point after injury, the standard deviations (SD) of the standard Ct values were calculated by SPSS. RESULTS: The expression trends of the seven genes were all found to be related to wound age, but there were lower variation coefficients and greater reliability of s TnI and Cox6c when compared with other genes. CONCLUSION The genes encoding structural proteins or proteins that perform basic functions can be suitable for wound age estimation.
Animals ; Contusions/genetics* ; Forensic Pathology ; Gene Expression Profiling ; Intercellular Adhesion Molecule-1 ; Muscle, Skeletal/metabolism* ; NF-kappa B ; Proteins ; RNA, Messenger/metabolism* ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Regression Analysis ; Reproducibility of Results ; Time Factors ; Wound Healing/genetics*

Cells in the body are exposed to physiological and pathophysiological stimuli that encompass both chemical and mechanical factors. It is important to understand how these factors modulate functions at cellular and organ levels. Compared to the large amount of information on cellular or organ responses to chemical factors, there is a paucity of knowledge on the effects of mechanical factors. Recent advances of fluorescence proteins and microscopy make it a very useful tool for elucidating the mechanotransduction processes; the state-of-the-art technologies for live-cell imaging of signaling is particularly valuable for investigating the spatial and temporal aspects of molecular mechanisms in mechanobiology. This review will cover the basic knowledge of fluorescence proteins and their application for biological research. In particular, the development and characterization of biosensors based on fluorescent resonance energy transfer (FRET) will be discussed. Genetically encoded FRET biosensors, which allows the imaging and quantification of tempo-spatial activation of molecules, will be introduced to demonstrate how the initiation and transmission of biochemical signals in response to local mechanical stimulation can be visualized in live cells. Specific emphasis will be on the elucidation of molecule hierarchy of signaling transduction in live cells upon the mechanical stimulation.

Objective To explore the associations between cesarean section with different types and intensity of physical activity in the second trimester pregnant women． Methods Six hundred and seventy－two participants from the Chinese pregnant women cohort study ( CPWCS) were analyzed． The pregnancy physical activity questionnaire ( PPAQ) was used to collect the status of physical activities in pregnant women． The participants were followed up and the data of delivery way was collected． Logistic regression model was conducted to analyze the associations． Ｒesults A total of 273 pregnant women ( 40. 63%) were delivered by cesarean section． After adjusting age，pre-pregnancy BMI and history of childbirth，results of Logistic regression model showed that pregnant women with higher levels of exercise had a lower risk of cesarean section than those who did not participate in exercise ( OＲ= 0. 564，95% CI: 0. 338－0. 941) . In terms of physical activity intensity，pregnant women who participated in the higher level of moderate to vigorous physical activity had a lower risk of cesarean section than those who partici- pated in the lower level ( OＲ= 0．642，95% CI: 0．437－0．972) ． Conclusions Exercise and moderate to vigorous physical activity are protective factors for cesarean section． Health education should be further strengthened to encourage pregnant women to carry out appropriate physical activity during pregnancy．

Self-microemulsifying drug delivery systems (SMEDDS) were developed to overcome the problems of delivery and administration of piroxicam, a drug with low bioavailability and gastrointestinal irritation, The in vitro properties of it were assessed. The solubility of piroxicam in several oils and surfactants was determined, and the compatibility of various oils and surfactants was investigated. Ternary phase diagrams were constructed to optimal area of microemulsion, and the influence of different oily phases, surfactants and co-surfactants was studied. The droplet size and dissolution of optimal formulation were determined to prove that the dosage form is a useful delivery system for piroxicam. In the optimized piroxicam SMEDDS, cinnamic alcohol was selected that gave the maximal solubility to piroxicam. Labrafil M 1944CS, Cremophor EL and Transcotol P were used as oils, surfactant and co-surfactant, respectively. Droplet size and distribution of three piroxicam SMEDDS formulations were (32.2 +/- 5.0), (40.1 +/- 6.4), (81.9 +/- 12.2) nm individually. And the releasing of piroxicam was rapid and complete. The optimized SMEDDS for piroxicam was obtained.
Administration, Oral ; Drug Compounding ; methods ; Drug Delivery Systems ; Emulsions ; Glycerides ; chemistry ; Glycerol ; analogs & derivatives ; chemistry ; Particle Size ; Piroxicam ; chemistry ; Polyethylene Glycols ; chemistry ; Propanols ; chemistry ; Solubility ; Solvents ; chemistry ; Surface-Active Agents ; chemistry

OBJECTIVE: To investigate the relationship between the expression of secreted frizzled-related protein 5 (SFRP5) mRNA and the time interval after skeletal muscle injury in rats by real-time PCR. METHODS: A total of ninety SD rats were randomly divided into the contusion groups at different times including 4h, 8h, 12h, 16h, 20h, 24h, 28h, 32h, 36h, 40h, 44h, 48h after contusion, incision groups at different times including 4h and 8h after incision and the control group. The samples were taken from the contused zone at different time points. The total RNA was isolated from the samples and reversely transcribed to analyze the expression levels of SFRP5 mRNA. RESULTS: Compared to the control group, the expression of SFRP5 mRNA in contusion groups were down-regulated within 48 h after contusion and reached the lowest level at 20 h, and the expression of SFRP5 mRNA gradually increased from 20 h to 48 h after contusion. The expression of SFRP5 mRNA in the incised groups were significantly lower than that of the contusion groups at 4 h after injury. At the time of 8 h, the expression levels between the contusion and incision groups showed no statistically significant difference. CONCLUSION It is suggested that SFRP5 mRNA analysis may show regular expression and can be a marker for estimation of skeletal muscle injury age.
Animals ; Biomarkers/metabolism* ; Contusions/metabolism* ; Membrane Proteins/metabolism* ; Muscle, Skeletal/metabolism* ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction

OBJECTIVETo classify the Chinese isolates of Coltiviruses.

METHODSThree sets of primers were selected among them two were specific to the 9th and 12th segments of subgroup B2, and one was for the 12th segment of subgroup B1-All the Chinese isolates of Coltivirus selected in the experiment were classified according to the lengths of different amplicons of the reverse transcriptase-polymerase Chain reaction (RT-PCR). The homogenicity of the nucleic acids of the isolates BJ95-75 and YN-6 was also compared with other Coltivirus strains belonging to subgroup B2.

RESULTSWith the primers 12-854-S/12-B2-R, which were specific to the 12th segment of Coltivirus subgroup B2-850 bp amplicons were obtained from Beijing isolate BJ95-75 and all the Yunnan isolates such as YN-6, -67-1, -68-1, -69, -70-1, -70-2, -90, -92-2, -93 of Coltivirus 492 bp DNA fragments were also amplified from all of them with the segment 9th specific primers 9-JKT-S/9-JKT-R. However no positive results were obtained from Northeast isolates NE97-12, NE97-31 and control viruses YN-99(Orbivirus),YN-151-1(JEV) with the same two sets of primers. With 12-B1-S/12-B1R primers specific to the 12th segment of subgroup B1, no amplicons of right length were obtained from any of the Chinese isolates of Coltivirus and the control viruses. When compared the nucleic acid sequences of BJ95-75 and YN-6 with other Coltivirus strains such as Bannavirus, JKT6423, JKT6969, JKT7043, the amplicons from segment 12th of these two strains had more than 89.4% homology with the other strains, especially to the earlier Chinese isolate Bannavirus, the homolog was more then 98.9%. Nearly 96.5% and 99.2% of the nucleic acids of the amplicons from segment 9th of the two strains were being homologous to Bannavirus and about 84.0% to JKT6423, which had been classified into type B2a. But the maximal homogenicity was about 53% when compared with the other two coltivirus strains. JKT6969 and JKT7043 which had been classified into type B2b.

CONCLUSIONGenotyping the recent Chinese isolates of coltivirus for the first time in our country. Most of the Chinese isolates belong to subgroup B2, more exactly type B2a. The Northeast isolates NE97-12 and NE97-31 were not correctly grouped with the available primers.

Animals ; Base Sequence ; China ; Coltivirus ; classification ; genetics ; isolation & purification ; Culicidae ; virology ; Genotype ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Homology, Nucleic Acid

This paper described the preparation and liver targeting traits of new solid lipid nanoparticles (SLN) containing floxuridinyl dibutyrate (FUDRB) modified with beta-D-galactosides (G2). FUDRB-SLN and FUDRB-G2SLN were prepared by thin layer ultrasonic technique. Transmission electron microscopy micrograph analysis demonstrated that the particle sizes of FUDRB-SLN and FUDRB-G2SLN were (137.5 +/- 11.1) nm and (95.0 +/- 10.7) nm. Drug loading were 9.64% and 8.56%, and entrapment efficiency were 99.81% and 96.23%, respectively. The concentrations of floxuridine (FUDR) in serum and some organs (liver, kidney and lung) were determined by RP-HPLC after iv administration of SLN. FUDR release was confirmed, and a significant enrichment of SLN modified with G2 was observed in liver with G2 complex (targeting rates of SLN-G2 was 8.28 for liver) in comparison with FUDR-sol (targeting rate was 2.56). FUDR could be detected in liver in mice at 480 min after iv administration of FUDRB-G2SLN. These results suggested that incorporation of G2 (4%-5%, g/g) into SLN enhanced the liver targeting-ability of FUDRB. SLN containing G2 could be a useful drug carrier system for liver targeting.
Animals ; Antimetabolites, Antineoplastic ; administration & dosage ; pharmacokinetics ; Area Under Curve ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Female ; Floxuridine ; administration & dosage ; blood ; pharmacokinetics ; Galactosides ; chemistry ; Lipids ; chemistry ; Liver ; metabolism ; Male ; Mice ; Nanoparticles ; Particle Size ; Tissue Distribution

OBJECTIVETo identify the molecular etiopathogenesis for a non-syndromic hearing loss patient.

METHODSThe core family, consists of the patient and his parents, was recruited. Genomic DNA was extracted from peripheral blood. Mutation analysis was carried out by SNaPshot and next-generation sequencing technology. Mutations in SLC26A4 gene were verified by polymerase chain reaction and direct sequencing.

RESULTSCompound heterozygous mutations p.V306GfsX24 and p.P516PfsX11 in SLC26A4 gene were detected in the patient, heterozygous mutation p.V306GfsX24 was detected in the father, heterozygous mutation p.P516PfsX11 was detected in the mother.

CONCLUSIONSCompound heterozygous mutations p.V306GfsX24 and p.P516PfsX11 contributed to patient's hearing loss. Next-generation sequencing technology is a useful tool for detecting de novo mutations of deafness genes, and is suitable for clinical application.

Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; DNA Mutational Analysis ; Deafness ; genetics ; Female ; Genetic Carrier Screening ; Genetic Testing ; Humans ; Male ; Membrane Transport Proteins ; genetics ; Mutation ; Pedigree

The authors describe here the procedures for using the gelatin half-embedding method to obtain thin spinal cord slices with attached dorsal roots and performing visually guided whole-cell patch-clamp recording of postsynaptic currents evoked by primary afferent fibers in rat spinal dorsal horn. A segment of spinal cord with attached dorsal roots was prepared and half-embedded in an agar block with 20% (w/v) gelatin. Thin spinal cord slices with attached dorsal roots were obtained with a vibratome and whole-cell patch-clamp configuration was established under the infrared observation. At the holding potential of -70 mV, spontaneous excitatory postsynaptic currents (EPSCs) and dorsal root stimulation-evoked EPSCs were recorded as inward currents. According to the conduction velocity of afferent fibers and stimulus threshold, evoked EPSCs that are mediated by A-like or C-like fibers were distinguished. At the holding potential of 0 mV, spontaneous inhibitory postsynaptic currents (IPSCs) and dorsal root stimulation-evoked IPSCs were recorded as outward currents. Using 5 micromol/L strychnine or 20 micromol/L bicuculline, GABAergic or glycinergic evoked IPSCs could be isolated. Using visual patch-clamp method synaptic transmission can be accurately assessed by measuring postsynaptic currents of the dorsal horn neurons. More importantly, with the aid of infrared observation, the incidence of failure to establish a clamp configuration can be greatly reduced and it becomes easier to make recordings from the neurons in deep dorsal horn laminae. Thus, the present research approach an effective approach to study the modulation of primary afferent synaptic transmission.
Animals ; Electrophysiology ; Evoked Potentials ; physiology ; Excitatory Postsynaptic Potentials ; physiology ; Female ; Male ; Neurons, Afferent ; physiology ; Patch-Clamp Techniques ; methods ; Posterior Horn Cells ; physiology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; cytology ; physiology ; Synaptic Transmission ; physiology