Aim To investigate the mechanism of dia-betes changing the hepatic CYP1 A2 through in vitro cell culture study.Methods The function of CYP1 A2 in HepG2 and Fa2N-4 cells were evaluated by determi-ning the level of phenacetin metabolism,and the mR-NA expression of CYP1 A2 in cells was detected by real time PCR.HepG2 cells were co-cultured with serum of diabetic rats(type 1 and type 2)and normal rats,then the CYP1 A2 function in cells were evaluated.Then, the HepG2 and Fa2N-4 cells were co-cultured with a series of concentrations of saturated (including palmitic acid and stearic acid)and unsaturated fatty acids(in-cluding oleic acid and linoleic acid)for 48 h,and the function and expression of CYP1 A2 in the cells were compared.Results It was found that the activities of CYP1 A2 were higher in cells incubated with diabetic serum of both type.All high concentration of fatty acids could increase the function and expression of CYP1 A2 in both HepG2 and Fa2N-4 cells.Conclusion It is speculated that the abnormal level of fatty acids under diabetic state might be part of the reasons why diabetes change the hepatic CYP1 A2,which provides the basis for future study.

Body fat of 12 male adults were measured by water displacement me-thod(density method) at every morning for 5 successive days. The standard deviation of single observation was 0.29kg calculated by mean residual lung volume method. It was significantly lower than the value (0.5kg) calculated by the ordinary method (p

OBJECTIVE: To explore the feasibility of detecting of Y-STR of fetal DNA in maternal plasma using Ion Torrent PGM™ System. METHODS: A total of 16 fetal DNA samples from maternal plasmas (8 cases from 38 weeks gestational age and 8 ones from 12 weeks) were prepared and a multiplex assay with 7 STR loci (DYS390, DYS391, DYS393, DYS438, DYS437, DYS456, DYS635) was designed for multiplex-PCR amplification. Using Ion Torrent PGM™ System, the results of Y-STR sequences and capillary electrophoresis were obtained and compared. RESULTS: Y-STR specific alleles were detected in the maternal plasma of all the pregnant women having male babies of second and third trimester, which were higher than that detected by capillary electrophoresis. Consistent Y-STR genotypes were observed between fetal DNA from maternal plasma and genomic DNA from the newborn babies. CONCLUSION Based on Ion Torrent PGM™ System, the prenatal Y-STR detection method may provide a high-sensitive and high-throughput choice for prenatal STR detection in forensic testing.
Alleles ; Chromosomes, Human, Y/genetics* ; DNA/blood* ; Family ; Female ; Fetal Blood/chemistry* ; Genotype ; Haplotypes ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Pregnancy ; Sensitivity and Specificity ; Sex Determination Analysis ; Tandem Repeat Sequences/genetics*

Objective To increase the transfection of EGFP-N3 plasmids into 293T cells using ultrasound-targeted microbubbles delivery(UTMD) mediated a peptide nucleic acid (PNA) binding nuclear localization signal (NLS).Methods Antibody-targeted microbubbles were used in the experiments which can specifically recognize the SV40T antigen receptor.The SV40T antigen receptors were expressed on the membranes of 293T cells.The PNA containing the NLS were inserted in the EGFP-N3 plasmid DNA,which increased nuclear localization.Ultrasound-targeted microbubble delivery (UTMD) and the PNA binding NLS were utilized to improve the cytoplasmic import of plasmids and the nuclear intake of the plasmid from the cytoplasm,respectively.The study was divided into five groups:Contrast (group A),Common microbubble + DNA (group B),Antibody-targeted microbubbles + DNA (group C),Common microbubbles + NLS-PNA-DNA (group D),Antibody-targeted microbubbles + NLS-PNA-DNA (group E).Fluorescence microscope was used to observe the fluorescent light in each group;flow cytometry to test the transfection;RT-PCR and Western blot to detect genes' mRNA and protein expression level.Results Ultrasound and antibody-targeted microbubble delivery (UTMD) significantly enhanced the cytoplasmic intake of exogenous genes and maintained high cell viability(＞80％).Fluorescent microscope showed that the quantities of green fluorescence in cells were increased successfully.The transfection results of flow cytometry were 0,(9.30 ± 0.46)％,(26.46 ± 2.01)％,(29.54 ± 0.62)％,(45.72 ± 1.86)％,respectively,and the differences were statistically significant(P ＜0.05).The relative mRNA and protein expression in group E were greater than those in group C and D respectively (P ＜0.05).Conclusions UTMD combined with antibody-targeted microbubbles and a PNA binding NLS plasmid can significantly improve transfection efficiency of exogenous genes by enhancing both cytoplasmic and nuclear DNA import.

Objective To analyze the clinical features and the correlation between radiographic parameters and incidence of calcaneal spur in the patients from Nanfang Hospital,Southern Medical University,China.Methods Three experienced observers independently used the image acquisition and transmission system (PACS) to collect the data of lateral and axial X-ray images of calcaneus or ankle joint in neutral position from the patients with calcaneal spur and normal controls who had undergone radiological examination in Nanfang Hospital,Southern Medical University from July 2014 through December 2015.Ten radiological parameters of the foot (B(o)hler angle,Gissane angle,calcaneal inclination angle,talocalcaneal angle,talus horizontal angle,posterior facet inclination angle,calcaneal length,height of the posterior facet,absolute foot height,and calcaneal width) were measured in both the patients and normal controls.The location,morphology and length of calcaneal spurs were compared between genders,sides and age groups.Results A total of 216 parpatients were included in the study.Female patients were more than male ones,simple plantar spurs more than simple achilles tendon ones,type B spurs more than type A ones,the length of achilles tendon spurs larger than that of plantar ones,female plantar spurs more than males ones,and the length of right foot plantar spurs larger than that of left foot ones.All the differences above were statistically significant (P ＜ 0.05).There were no significant differences in the location,morphology or length of calcaneal spurs between the age group of ≤ 60 years old and the age group of ＞ 60 years old (P ＞ 0.05).The incidence of calcaneal spur were significantly correlated to Gissane angle (P =0.000,OR =0.944,95％ CI 0.917-0.973),posterior facet inclination angle (P=0.017,OR=0.957,95％ CI 0.924-0.992) and height of the posterior facet (P =0.007,OR =0.933,95％ CI O.886-0.981).Conclusions Calcaneal spur favored more females than males.Plantar spurs were more common than Achilles ones.Plantar spurs of Type B were more common than those of Type A.Achilles spurs were longer than plantar ones.More females suffered plantar spur than males.Right foot spurs were longer than left foot ones.Age had no significant influence on the spur characteristics.The incidence of spur might have been related to the Gissane angle,posterior facet inclination angle and height of the posterior facet of the foot.

Objective:To explore the monitoring value of left ventricular functional parameters obtained by bedside ultrasound combined with clinically relevant indicators in patients with veno-arterial extracorporeal membrane oxygenation (VA-ECMO).Methods:A retrospective study was conducted. A total of 24 patients receiving VA-ECMO adjuvant support in Renmin Hospital of Wuhan University from June 2018 to January 2020 were selected. The bedside ultrasound was performed on the first day of ECMO support, the day before weaning, the clinical indicators before weaning were obtained. The differences in clinical indicators and the left ventricular functional parameters between the two groups of whether weaning successfully were compared; univariate Logistic regression analysis was used to screen out the related factors affecting weaning.Results:Sixteen patients were successful weaned and 8 patients failed. Compared with the weaning failure group, patients in the weaning success group required less continuous renal replacement therapy (CRRT, cases: 4 vs. 6, P < 0.05), mean arterial pressure (MAP) before weaning was higher [mmHg (1 mmHg = 0.133 kPa): 84.64±9.55 vs. 62.30±8.79, P < 0.05], and the pulse oxygen saturation (SpO 2) was also higher (0.966±0.670 vs. 0.866±0.061, P < 0.05), while vasoactive-inotropic score (VIS), serum creatinine (SCr) and serum lactic acid (Lac) were lower [VIS score: 7.27±1.42 vs. 16.93±8.52, SCr (μmol/L): 123.60±83.64 vs. 213.10±117.39, Lac (mmol/L): 1.94±0.91 vs. 5.62±5.48, all P < 0.05]. Univariate Logistic regression analysis showed that the MAP, VIS, SCr, Lac, SpO 2 before weaning were the related factors affecting weaning [odds ratio ( OR) were 0.306, -0.740, -0.011, -0.632, -4.069; 95% confidence interval (95% CI) were 1.065-1.732, 0.235-0.899, 0.979-0.999, 0.285-0.992 and 0.001-0.208; P values were 0.014, 0.022, 0.038, 0.047, 0.002]. In the weaning success group, left ventricular ejection fraction (LVEF), velocity of mitralannulus in systolic (LatSa), maximum flow velocity of aortic valve (AV-Vmax), velocity-time integral (VTI), left ventricular global longitudinal strain (LVGLS), left ventricular global longitudinal strain rate (LVGLSr) were all increased on the day before ECMO weaning compared with the first day of ECMO support [LVEF: 0.40±0.05 vs. 0.28±0.07, LatSa (cm/s): 6.81±0.91 vs. 4.62±1.02, AV-Vmax (cm/s): 104.81±33.98 vs. 64.44±16.85, VTI (cm): 14.56±3.11 vs. 7.96±1.98, LVGLS: (-8.95±2.59)% vs. (-5.26±1.28)%, LVGLSr (1/s): -0.48±0.11 vs. -0.29±0.09], whereas the ECMO flow was significantly reduced (L/min: 1.46±0.47 vs. 2.64±0.31), the differences were statistically significant (all P < 0.05). There was no significant difference in left ventricular functional parameters between the first day of ECMO support and the day before ECMO weaning in the weaning failure group. Compared with the weaning failure group, the weaning success group had higher LVEF, LatSa, AV-Vmax, VTI, LVGLS, LVGLSr on the day before ECMO weaning [LVEF: 0.40±0.05 vs. 0.26±0.07, LatSa (cm/s): 6.81±0.91 vs. 4.31±1.03, AV-Vmax (cm/s): 104.81±33.98 vs. 67.67±18.46, VTI (cm): 14.56±3.11 vs. 7.75±2.77, LVGLS: (-8.95±2.59)% vs. (-4.81±1.81)%, LVGLSr (1/s): -0.48±0.11 vs. -0.30±0.10, all P < 0.05] and lower ECMO flow (L/min: 1.46±0.47 vs. 2.20±0.62, P < 0.05). Conclusion:Bedside echocardiographic left ventricular function parameters (LVEF, LatSa, AV-Vmax, VTI, LVGLS, LVGLSr) combined with clinical indicators (MAP, VIS, SCr, Lac, SpO 2) were helpful to evaluate the therapeutic effect of patients receiving VA-ECMO support and can provide important guiding value in the selection of VA-ECMO weaning timing and the judgment of prognosis.

Objective To explore the effects of amniotic lacrimal duct stenting on the prevention of dry eye in castrated rabbits.Methods Thirtysix healthy male rabbits were selected,the third eyelid were cut off and antiinfection treatment were given,which were randomly divided into 3 groups (12 cases in each group),the castrated male rabbits models were made.Among them,group A was negative control group,group B was dry eye model group,group C was group of lacrimal amniotic membrane group.At 2 weeks before implantation of amniotic lacrimal duct stent,2 weeks,4 weeks and 6 weeks after implantation,the fluorescent (FL) examination,Western blot,Schirmer I examination,immunofluorescence staining and corneal confocal microscopy were performed.Results The levels of tear secretion and FL in the three groups among different time points were significantly different (F=7.126,P =0.009;F =9.658,P =0.016),and there were significant differences among three groups (F =12.582,P =0.005;F =13.187,P =0.013).The tendency of tear secretion and FL in the three groups were also significantly changed (F =8.531,P =0.007;F =10.652,P =0.019).The epithelial basal cells at 6 weeks after implantation in three groups were 3811 ±414,3820 ± 314,2789 ± 353,and the density of inflammatory cells was 266 ±28,266 ± 29,67 ± 13,there were significant differences among three groups (F =13.442,P =0.012;F =9.231,P =0.021).The K1 6 staining in the duct epithelium were negative,and the expression of α-SMA in the lacrimal duct tissue of group A,B and C was not changed at all time points after implantation of amniotic lacrimal stent,and there was no significant difference (F =14.681,P =0.002).Conclusion The amniotic lacrimal stent implantation has certain effect on the prevention of dry eye in rabbit.

Objective To investigate the protective effect and the underlying mechanism of water soluble coenzyme Q10 (CoQ10)against rotenone induced injury on PC12 cells model.Methods PC12 cells were cultured with rotenone,water-soluble CoQ1 0 was added to the culture media 3 hours prior to the rotenone incubation.We determined cell viability by CCK8;reactive oxygen species (ROS)was detected by spectrophotometer;and Bcl-2, Bax,active Caspase-3,Caspase-9 and apoptosis-inducing factor (AIF)were measured by Western blotting after 24-hour rotenone incubation.Results After the treatment by rotenone,cell viability decreased significantly (P＜0.01)and ROS level increased (P＜0.01).CoQ10 could improve PC12 cell viability (P＜0.01)and reduce the level of ROS (P＜0.01).Western blotting experiments showed that CoQ10 could reduce rotenone-induced Caspase-9 (P＜0.05),active Caspase-3 (P＜0.05)and Bax (P＜0.01)expressions,increase the expression of Bcl-2 (P＜0.01),and prevent nuclear translocation of AIF (P＜0.05).Conclusion CoQ10 has a protective effect on rotenone-induced apoptosis in PC12 cells,the mechanism of which may be through scavenging ROS in cells;decreasing caspase-9 ,active caspase-3 and Bax expressions;and increasing the expression of Bcl-2 ;and preventing AIF nuclear translocation.

OBJECTIVETo discuss our experience on the diagnosis and treatment of thoracic aorta rupture (TAR) that is one of the main common causes of death in the victims under blunt chest trauma.

METHODSBetween July 2001 and March 2006, 9 patients (6 men and 3 women, aged from 20 to 54 years) suffering from acute traumatic aorta rupture after motor vehicle accidents received emergent surgical treatments in our hospital. Based on our experience in the rescue of the first TAR patient we introduced a practical procedure on the diagnosis and treatment of TAR in our department. All the other patients generally followed this procedure. Eight patients received contrast material enhanced helical computerized tomography scan before the operation. The leakage of constrast medium from the aorta isthmus was found, and diagnosis of TAR was confirmed. Seven patients underwent immediate operation within 14 hours after accidents. One patient was treated on the 5th day of the accident because of delayed diagnosis of aortic rupture. All patients received general anesthesia with double lumen endotracheal tube and normothermic femoro-femoral partial cardiopulmonary bypass, with beating heart and aortic clamping. One patient received simple repair, and others received partial replacement of thoracic aorta with artificial vascular graft.

RESULTSSeven TAR patients were successfully salvaged. Three patients combined brain injury as well as extremitiy hemiplegia before operation. After treatments one was fully and two partially recovered without paraplegia.

CONCLUSIONSProper practical protocol is emphasized for the surgical repair of TAR because it will reduce the mortality of severe blunt chest injury.

Accidents, Traffic ; Adult ; Aorta, Thoracic ; injuries ; Aortic Rupture ; diagnosis ; surgery ; Emergencies ; Female ; Humans ; Male ; Middle Aged ; Rupture

OBJECTIVETo clone migfilin-N terminal sequence into E.coli and obtain a fusion protein for preparing rabbit polyclonal antibody against migfilin, thereby facilitating the study of the role of migfilin in the biological behavior of colon cancer.

METHODSBased on human migfilin cDNA sequence, a pair of primers was designed to amplify migfilin-N terminal sequence by PCR. The PCR product was subcloned into the bacterial expression vector pGEX-4T-1 with EcoRI/XhoI sites, and the target recombinant plasmids were identified with enzymatic cleavage followed by DNA sequence analysis. By transforming the expression vector into component E.coli BL(21) cells, the GST-migfilin-N fusion protein was expressed with IPTG induction. Glutathione-sepharose beads were used to purify the fusion protein, and anti-migfilin polyclonal antibody was produced by immunization of rabbits with the purified GST-migfilin N-terminal fusion protein. The resultant anti-migfilin polyclonal antibody was purified by protein A beads and used for Western blotting for detecting migfilin expression in different cell lines.

RESULTS AND CONCLUSIONThe migfilin-N terminal gene fragment was cloned successfully, and purified GST-migfilin N-terminal fusion protein and anti-rabbit migfilin polyclonal antibodies were obtained. Western blot analysis demonstrates that the antibodies specifically detected migfilin expression in the cell lines, which may facilitate further investigation of the role of migfilin in the biology of colon cancer.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; isolation & purification ; Base Sequence ; Blotting, Western ; Cell Adhesion Molecules ; genetics ; immunology ; Cell Line, Tumor ; Cloning, Molecular ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; Cytoskeletal Proteins ; genetics ; immunology ; DNA, Complementary ; chemistry ; genetics ; Escherichia coli ; genetics ; Humans ; Molecular Sequence Data ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA