Efficiency of preeclampsia screening based on biomarkers of aneuploidy screening in first trimester / 中华围产医学杂志

Objective:To evaluate the performance of biomarkers in aneuploidy screening in the first trimester-pregnancy associated plasma protein A(PAPP-A) combined with Fetal Medicine Foundation (FMF)'s competing risk model in screening preeclampsia among our population.Methods:This study was based on a prospective cohort of singleton pregnant women who underwent aneuploidy screening in the first trimester in Nanjing Drum Tower Hospital from January 2017 to September 2020. Mean arterial pressure (MAP), uterine artery pulsatility index (UtA-PI), and PAPP-A were converted into multiples of median (MoM) using the algorithm disclosed on the website of the FMF (fetalmedicine.org). The predictive outcomes of maternal factors alone or in combination with MAP, UtA-PI, and PAPP-A (alone or in combination) were calculated. Chi-square test, Fisher's exact test or rank sum test were used for comparison among groups and Bonferroni method for pairwise comparisons. Receiver operating characteristic (ROC) curve was used to evaluate the screening efficiency and to calculate the sensitivities of predicting preeclampsia, term and preterm preeclampsia at false-positive rates of 5% and 10%. The predictive performance of this model was further compared to the screening strategy that was recommended in Diagnosis and treatment of hypertension and pre-eclampsia in pregnancy: a clinical practice guideline in China (2020). Results:Among the 5 144 singleton pregnancy women who were recruited in the cohort, 4 919 cases were included and analyzed in this study. A total of 223 cases were diagnosed as preeclampsia (4.5%), including 55 preterm (1.1%) and 168 term preeclampsia (3.4%). The median of MoM values of MAP, UtA-PI, and PAPP-A in the non-preeclampsia group were around 1.0±0.1. Statistical significance was observed in the difference of MAP, UtA-PI, and PAPP-A Mom between women with preterm preeclampsia and those without preeclampsia [1.061 (0.999-1.150) vs 0.985 (0.935-4.043), 1.115 (0.873-1.432) vs 1.039 (0.864-1.236), 0.820 (0.493-1.066) vs 1.078 (0.756-1.508)], which was also seen in the difference of MAP and PAPP-A Mom between women with term preeclampsia and those without preeclampsia [1.065 (1.002-1.133) vs 0.985 (0.935-4.043), 1.007 (0.624-1.393) vs 1.078 (0.756-1.508)] (all P<0.025). The combination screening with maternal factors+MAP+UtA-PI+PAPP-A was noted for the best efficiency. In predicting preeclampsia preterm and term preeclampsia at the false-positive rate of 10%, the sensitivity of the model was 53.0%, 76.4% and 44.6% respectively. Using the screening method recommended in Diagnosis and treatment of hypertension and pre-eclampsia in pregnancy: a clinical practice guideline in China(2020), the proportion of people at high risk of preeclampsia was 5.9% (290/4 919), and the sensitivity for predicting preterm preeclampsia was 25.5% (14/55), which was significantly lower than the combination screening with maternal factors+MAP+UtA-PI+PAPP-A [65.5% (36/55)] when using the same proportion of high-risk population. Conclusion:The preeclampsia screening model based on aneuploidy screening biomarkers in the first trimester--PAPP-A in combination with materral factors, MAP, UtA-PI, can effectively screen preterm preeclampsia in the local population without increasing the laboratory costs.

Estrogen Promotes cAMP Production in Mesenchymal Stem Cells by Regulating ADCY2

Background and Objectives@#The maternal-fetal interface is an important source of mesenchymal stem cells (MSCs), and it is influenced by high levels of estradiol (E2) during pregnancy. It is highly important to study the role of E2 in MSCs for both clinical application and understanding of the mechanisms underlying pregnancy related diseases. @*Methods@#and Results: In this study, differently expressed genes (DEGs) were found in the MSCs after exposure to E2. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs was performed and the integrated regulatory network of DEGs-miRNA was constructed. A total of 390 DEGs were found in the MSCs exposed to E2, including 164 upregulated DEGs (e.g. ADCY2, VEGFA and PPY) and 226 downregulated DEGs (e.g. KNG1, AGT and NPY). Additionally, 10 miRNAs (such as miR-148A/B, miR-152, miR-182) identified the integrated regulatory network of DEGs-miRNAs. Among them, the expression of ADCY2 was significantly upregulated, and this was associated with multiple changed genes. We confirmed that the expression of ADCY2 is significantly promoted by E2 and subsequently promoted the production of cAMP in MSCs. We also found that E2 promoted ADCY2 expression by inhibiting miR-152 and miR-148a. @*Conclusions@#E2 promotes the expression of cAMP through miR-148a/152-ADCY2 in MSCs. It is suggested that E2 plays a key role in the growth and function of MSCs.

Clinical research of Qingfei-Xiaoyan Decoction combined with conventional western medicine in patients with acute exacerbation of chronic obstructive pulmonary disease / 国际中医中药杂志

Objective:To observe the clinical efficacy of Qingfei-Xiaoyan Decoction combined with conventional western medicine in patients with acute exacerbation of chronic obstructive pulmonary disease (COPD). Methods:Eighty-two patients with acute exacerbation of COPD were selected in Bozhou People's Hospital from January 2017 to June 2018 and randomly divided into a control group and a treatment group (41 in each group) using randomized number table method. Patients in the control group were treated with conventional western medicine and those in the treatment group with Qingfei-Xiaoyan Decoction on the basis of the control group for 2 weeks. Dyspnea of patients was evaluated with the modified British Medical Research Council dyspnea questionnaire (mMRC). Impact of the disease was measured with the COPD Assessment Test (CAT). The serum levels of C-reactive protein (CRP) and procalcitonin (PCT) were measured by ELISA. Results:The total efficacy rate in trementat group 95.1% (39/41) was significantly higher than that in the control group 73.2% (30/41) ( χ2=4.999, P=0.025). After the treatment, the scores of mMRC (0.63 ± 0.07 vs. 0.95 ± 0.12; t=7.921, P<0.01) and CAT (9.18 ± 0.12 vs. 14.01 ± 1.56; t=11.359, P<0.01) in the treatment group were significantly lower than those in the control group. After the treatment, the forced expiratory volume in one second (FEV1) percentage predicted (51.05% ± 5.63% vs. 45.77% ± 5.31%; t=10.453, P<0.01) and FEV1/forced vital capacity (FVC) (59.15 ± 6.44 vs. 54.24 ± 6.02; t=5.621, P<0.01) in the treatment group were significantly higher than those in the control group. After the treatment, the serum levels of CRP (8.06 ± 0.87 mg/L vs. 10.55 ± 1.21 mg/L; t=10.216, P<0.01) and PCT (4.20 ± 0.48 μg/L vs. 6.33 ± 0.69 μg/L; t=7.004, P<0.01) in the treatment group were significantly lower than those in the control group. Conclusions:Qingfei-Xiaoyan Decoction can inhibite inflammation, improve symptoms and lung function, increase the efficacy in patients with acute exacerbation of COPD.

Effects and potential mechanisms of bradykinin B2 receptor on growth and biological function of human extravillous trophoblast cells / 中华围产医学杂志

Objective To investigate the effects and its mechanisms of bradykinin B2 receptor (B2R) on the growth and function of human extravillous trophoblast cells (HTR-8/SVneo cells).Methods B2R expression plasmid (pcDNA3.1-B2R) was constructed and B2R-specific small interfering RNA (siRNA) was synthesized.HTR-8/SVneo cells were divided into four groups and transfected with pcDNA-3.1 (blank plasmid group),pcDNA3.1-B2R (B2R expression plasmid group),siRNA negative control and B2R-specific siRNA,respectively.Quantitative real-time reverse transcription-polymerase chain reaction and Western blot were used to detect the changes in the expression of B2R,matrix metalloproteinase-2,matrix metalloproteinase-9,cyclin D1 and vascular endothelial growth factor-A at both mRNA and protein levels in HTR-8/SVneo cells.Cell counting kit-8 and flow cytometry were used to detect cell activity and cell cycle,respectively.Cell migration assay and cell invasion assay were used to detect cell migration and invasion,respectively.Tube formation assay was used to evaluate the tube formation abilities of HTR-8/SVneo cells.All data were analyzed with t test.Results (1) Compared with the blank plasmid group,expression of B2R in HTR-8/SVneo cells in the B2R expression plasmid group were significantly increased at both mRNA (5.06±0.49 vs 1.00±0.28,t=7.226,P=0.002) and protein levels (1.34 ± 0.07 vs 1.00± 0.05,t=3.727,P=0.006).And the expression of B2R in HTR 8/SVneo cells transfected with B2R-specific siRNA were significantly reduced at both mRNA (0.34±0.05 vs 1.00±0.17,t=3.667,P=0.021) and protein levels (0.74±0.03 vs 1.00±0.05,t=4.097,P=0.006) comparing with the siRNA negative control group.(2) Compared with the blank plasmid group,HTR-8/SVneo cells being transfected with B2R expression plasmid showed a higher proliferation activity (1.50 ±0.03 vs 1.34± 0.04) promoting G0/G1 to S phase transition;compared with the siRNA negative control group,B2R-specific siRNA inhibited the proliferation of HTR-8/SVneo cells (1.06 ± 0.04 vs 1.20± 0.02) and arrested the cell cycle at G0/G 1 phase (all P＜0.05).(3) Compared with the blank plasmid group,B2R expression plasmid significantly increased the HTR-8/SVneo cell migration distance [(80.67±0.33) vs (41.33±5.24) μm],the number of cells penetrating matrigel gel (360.70 ±12.33 vs 268.70 ±14.45) and the number of cells having tube-like structures (28.20 ± 2.47 vs 14.00± 1.67),while significantly decrease was shown in these three parameters in B2R-specific siRNA group comparing with the siRNA negative control group [HTR-8/SVneo cell migration distance:(56.00±3.51) vs (87.00±1.53) μ m,number of cells penetrating matrigel gel:143.30± 12.91 vs 252.30± 17.07;number of tube-like structures:6.25±1.49 vs 15.75 ±2.02;all P＜0.05].(4) Expression of matrix metalloproteinase-2 and matrix metalloproteinase-9 at mRNA level,and expression of cyclin D1 and vascular endothelial growth factor-A increased in the B2R expression plasmid group than in the blank plasmid group,and decreased in the B2R-specific siRNA group than in the siRNA negative control group at both mRNA and protein levels (all P＜0.05).Conclusions B2R might enhance the activity,migration,invasion and tube formation ability of human extravillous trophoblast cells through promoting the expression of matrix metalloproteinase-2,matrix metalloproteinase-9,cyclin D1 and vascular endothelial growth factor-A.

Pravastatin inhibits microRNA-155 expression and improves functions of lipopolysaccharide-treated human extravillous trophoblast cells / 中华围产医学杂志

Objective To investigate the effects ofpravastatin on the expression ofmicroRNA-155 (miR-155) and the functions of lipopolysaccharide (LPS)-treated extravillous trophoblast cells.Methods In vitro cultured HTR-8/SVneo cells were divided into the following groups:control group,enhanced plasmid with green fluoscent protein (pEGFP)-miR-155 group (transfected with green fluorescent protein-tagged miR-155),LPS group (treated with 100 ng/mL of LPS),miR-155 inhibitor+LPS group,pravastatin+LPS group (treated with 100 ng/mL of LPS following pretreatment with 12.50,25.00,50.00 and 100.00 μ g/ml of pravastatin),and pravastatin+pEGFP-miR-155 group (transfected with pEGFP-miR-155 following pretreatment with 50 μ g/ml of pravastatin).Levels of miR-155 in HTR-8/SVneo cells treated with different strategies were measured by real-time polymerase chain reaction.Expression of phosphorylated JunB (p-JunB) and p-FosB proteins was analyzed by Western blotting.Migration,invasion and apoptosis of HTR-8/SVneo cells were also analyzed.All data were analyzed with t test.Results (1) Compared with the control group,HTR-8/SVneo cells in the pEGFP-miR-155 group were characterized by shorter migration distance [(274.70± 18.82) vs (181.00±8.62) μ m],less transmembrane cells [(123.00±4.36) vs (63.00±6.08)] and enhanced apoptosis [(5.40± 0.68)％ vs (9.27±0.68)％] (all P＜0.05).(2) Compared with the LPS group,the miR-155 inhibitor+LPS group showed longer migration distance of HTR-8/SVneo cells [(166.30±5.07) vs (242.00±18.07) μm,P＜0.05],more transmembrane cells [(71.67±6.12) vs (109.00±7.81),P＜0.05] and decreased cell apoptosis [(14.40±1.69)％ vs (6.23± 0.44)％,P＜0.05].(3) Expression of miR-155 at mRNA level in the LPS group was increased as compared with that of the control group (1.65 0.07 vs 0.79±0.12,P＜0.05).Compared with the LPS group,pretreatment with 12.50,25.00,50.00 and 100.00 μ g/ml of pravastatin decreased the expression of miR-155 at mRNA level [(1.14±0.10),(1.02±0.10),(0.74±0.15) and (1.140.02)],especially at the concentration of 50 μμ g/ml (all P＜0.05).(4) Expression ofp-JunB and p-FosB proteins in the control,LPS and pravastatin (50 μ g/ml)+LPS groups were (0.33 ±0.06) vs (0.37±0.07),(1.22±0.20) vs (0.80±-0.13),and (0.31 ±0.02) vs (0.21 ±0.05),respectively,showing higher expression level in both p-JunB and p-FosB proteins in the LPS group compared with that of the other two groups (all P＜0.05).(5) Compared with the LPS group,the pravastatin (50 μμ g/ml)+LPS group showed increased migration distance [(166.30±5.07) vs (246.80± 13.42) μ m,P＜0.05],increased numbers of transmembrane cells [(71.67 ± 6.12) vs (95.33 ± 2.73),P＜0.05] and decreased cell apoptosis [(14.40± 1.69) vs (6.05 ± 0.35)％,P＜0.05].(6) Compared with the pEGFP-miR-155 group,the pravastatin (50.00.00 μμ g/mL)+pEGFP-miR-155 group showed longer migration distance [(197.50± 13.86) vs (275.80± 13.63) μ m,P＜0.05],more transmembrane cells [(52.67±5.49) vs (125.00±6.66),P＜0.05] and lower rate of cell apoptosis [(8.90± 1.00) vs (5.05±0.35)％,P＜0.05].Conclusions Pretreatment of extravillous trophoblast cells with pravastatin can protect them from apoptosis and loss of migratory and invasive abilities through inhibiting the activation of AP-1 and down-regulating the expression of miR-155,which may be a mechanism that inhibits the development of preeclampsia.

Optimizing the strategy of radiological examination reduces iatrogenic radiation exposure in major trauma patients / 中华创伤杂志

Objective To investigate the effect of optimized radiological examination strategy on iatrogenic radiation exposure in severe trauma patients so as to provide scientific basis for standardized application of radiological examination.Methods A controlled, three-stage intervention study from April 2010 to November 2011 was carried out.From April 2010 to July 2010, a pre-intervention study was conducted and enrolled 60 patients [43 males, 17 females;age (50 ± 14)years, age range 23-78 years].From August 2010 to March 2011, optimized strategies of radiological examination were implemented, including improving clinicians' knowledge to the standardization of radiological examination and iatrogenic radiation injury and limiting frequency of CT scans through the electronic medical record.From April 2011 to November 2011, post-intervention study was conducted and enrolled 100 patients (81 males, 19 females;age (47 ± 14) years, age range 18-79 years].During this period, major trauma patients were analyzed with respect to the clinical information, radiation examination frequency, ionizing radiation dose and influencing factors.Radiation examination frequency and radiation dose were compared before and after the intervention.Results Radiological examinations were mainly X-ray and CT before the implication of optimized strategies.Of the 60 patients, median frequency of X-rays and CT scan was 6.0(3.0-11.0) and 10.0(8.0-13.8).Median frequency of CT scan was positively correlated with the injury severity score (ISS) and ICU length of stay (r =0.369 and 0.523, P ＜ 0.05).Of the 100 patients, median frequency of CT scan was significantly reduced after the optimization of radiological examination (8.0 vs.10.0, P ＜ 0.05).Total frequency of radiological examination was significantly reduced as well (13.6 vs.17.8, P ＜0.01).There was no significant difference in the treatment success rate before and after the optimization of radiological examination (85.0％ vs.88.3％, P ＞ 0.05).When the frequency of head and chest CT scan was limited, the frequency of radiological examination, radiation exposure and radiological examination expenses were greatly reduced.Conclusions Too much X-ray,CT or other radiological examinations are noted in major trauma patients during the treatment period.Improved understanding of radiation-induced injury, optimizing radiological examination and controlling the repeated radiological examinations of the same site contribute to reducing iatrogenic radiology exposure without affecting the outcome.

Phenotypic and genetic analysis of a child with blepharophimosis, ptosis, epicanthus inverses syndrome and tetralogy of Fallot / 中华医学遗传学杂志

OBJECTIVE To determine the genetic cause of a child with blepharophimosis, ptosis, and epicanthus inverses syndrome and tetralogy of Fallot, and to correlate the phenotype with the genotype. METHODS Routine G-banding has been previously performed on the patient and her parents. Chromosome microarray analysis (CMA) was performed for the three individuals and the fetus. RESULTS Chromosomal analysis has suggested normal karyotypes for the child and her parents. However, a de novo 8.9 Mb deletion on chromosome 3q22.1-q23 was detected by CMA. The deleted region has encompassed 74 genes including 41 disease-related genes, and this is also the most frequent region involved in interstitial 3q deletion. Patients with deletion of this region often have a common feature of dysplasia of eyelids, as well as a spectrum of other anomalies according to different breakpoints, including microcephaly, skeletal anomalies, congenital heart defects, cranial anomalies, intellectual disability and developmental delay. The patient's phenotype was in accordance with such spectrum. Her parents and sib did not show this variation by CMA. CONCLUSION The de novo interstitial deletion of 3q22.1-q23 probably underlies the main clinical manifestation in this child. CMA can provide more detailed information and allow further investigation of the genotype-phenotype correlation.

Effects of tube size for drainage on flow rate of different kinds of fluid studied in vitro / 中华急诊医学杂志

Objective To determine the appropriate size of the tube for the thoracic drainage in good efficiency by the experimental study in the influence of the tube size on the flow rate of the fluid with different properties.Methods Three groups were divided according to the different components in the fluid:group A,whole blood with 30％ hematocrit; group B,2.5％ albumin solution; and group C,0.9％ normal saline.The total volume of the fluid was 1000 mL in each group in the experiment.Different sorts of fluids were drained with the chest tubes with different diameters (6F,8F,10F,12F,14F,16F,18F,20F,22F,24F,26F,28F,30F,32F,34F,36F of French F) separately,and the flow rate was calculated.ANOVA was used for the comparison of the differences in flow rate among the groups with given fluid property.Twofactor analysis of variance was used for the analysis of flow rates of fluid with different fluid properties.Curve fitting was performed according to the Poiseuille formula.Results The flow rate was positively correlated with the size of the chest drainage tube.The difference in flow rate among the tubes with difference in size was statistically significant (P ＜ 0.05) but there was no noticeable difference in flow rate between 6F and 8F (P =0.513).The flow rate of the 6F and 8F tubes was higher than that of the control (3.33 mL/min) but there was no significant difference between them (P ＞ 0.05).The flow rate of the tubes in 10F and above was obviously higher than that of control (P ＜ 0.05).The curve was estimated that group A was Q =0.002 9x4,R2 =0.991; group B Q=0.003 2x4,R2 =0.981; group C Q =0.003 4x4,R2 =0.975.When the flow rate was fixed at 3.33 mL/min,the estimated curve in group A was X ≈ 5.82F.Conclusions Our experiment indicated that the chest tube with small diameters (6F-14F) could meet the demand of high efficient drainage in the patients with hemothorax or pleural effusion.

Mesenchymal stem cell-conditioned medium could ameliorated triptolide induced damage in KGN cells / 中国免疫学杂志

Objective:To explore the ameliorative effect and machanism of MSCs conditioned medium on the ovarian granulosa cells damage induced by triptolide.Methods: Cell Counting Kit 8 assay was used to examine the cell vitality of KGNs with the treatment of triptolide.The mixed enzyme digestion method were used for the isolation of human umbilical cord mesenchymal stem cells (MSCs),and flow cytometry was used for the subsequent immunotype identification.MSCs conditioned medium was collected ,and Cell Counting Kit 8 assay and PI staining was used to analyse the effect of MSCs conditioned medium on the cell vitality and cell cycle distri -bution of triptolide-damaged KGN.Real-time PCR method was used to examine the expression of cell cycle related gene CDKN1A.Results:Triptolide can inhibit KGN cell growth with the inhibition of cell vitality and cell cycle of KGN.MSCs conditioned medium did not influence the proliferation and cell cycle of normal KGN , but improved the triptolide-induced vitality inhibition and extent of S-phase arrest, and inhibit the abnormal up-regulation of CDKN1A in KGN.Conclusion: MSCs conditioned medium ameliorated the KGN cell damage induced by triptolide.

Effects of perioperative enteral nutrition on host immunologic function in elderly patients with gastrointestinal tract malignant tumor / 中国医师进修杂志

Objective To evaluate the effects ofperioperative enteral nutrition on host immunologic function in elderly patients with gastrointestinal tract malignant tumor. Methods Eighty elderly patients with gastrointestinal tract malignant tumor were divided into two groups by random digits table with 40 cases each: experimental group was given enteral nutrition and control group was given parenteral nutrition.Peripheral blood samples were collected at the fifth and the first day before operation, and the fust and the seventh day after operation to examine the NK cell,T-lymphocyte subsets (CD3, CD4,CD8) and immunoglobulin (lgG, lg A, IgM ). Result There were significantly increased in NK cell [(40.1 ± 4.9)%], CD3[(69.40 ±7.10)%], CD4[(39.20 ± 5.83)%], IgA[(2.94 ±0.56) g/L] and IgM[(1.34 ±0.72) g/L] of experimental group compared with those of control group [(26.4 ± 4.9 )%, ( 61 .40 ± 8.32)%, ( 29.20 ± 5.65 )%, ( 1.72 ±0.76) g/L, (0.97 ± 0.89 ) g/L] (P ＜ 0.05 ). Conclusion The application of enteral nutrition in perioperative period can improve the immunologic function of elderly patients with gastrointestinal tract malignant tumor,reduce surgical risk and postoperative complications.

Glossy ganoderma spore oil promotes apoptosis of human lung adenocarcinoma SPC-A1 through downregulation of miR-21 / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effects of glossy ganoderma spore oil on the proliferation, apoptosis, expression of miR-21 and its target genes of human lung adenocarcinoma SPC-A1 cell line, and to explore its possible mechanism.</p><p><b>METHOD</b>The SPC-A1 cells were treated with glossy ganoderma spore oil for 24 and 48 hours. The inhibition growth efficacy was determined using cell count kit (CCK-8). Cell morphological changes were observed by light microscopy. Cell apoptosis was analyzed by flow cytometry. The expression of miR-21, PTEN and PDCD4 were determined by Real-time PCR.</p><p><b>RESULT</b>Glossy ganoderma spore oil concentration-dependently inhibited the SPC-A1 cell's proliferation. When the concentration of glossy ganoderma spore oil attained to 0.2%, the cells' morphology changed obviously. Glossy ganoderma spore oil could induce the apoptosis of SPC-A1 cells at low concentration. Glossy ganoderma spore oil down-regulated the expression of miR-21 and up-regulated the expression of PTEN and PDCD4 significantly.</p><p><b>CONCLUSION</b>glossy ganoderma spore oil could inhibit the proliferation obviously and cause the changes of cell morphology. Furthermore, glossy ganoderma spore oil induced apoptosis of SPC-A1 cell through down-regulating the expression of miR-21 and up-regulating tumor suppressors.</p>

Effect of Moschus Combined with Borneol on Brain Water Content and Blood-Brain Barrier Permeability in Rat Model of Cerebral Focal Ischemia with Reperfusion / 广州中医药大学学报

【Objective】To observe the effect of Moschus combined with Borneol on brain water content and blood-brain barrier(BBB)permeability in rat model of cerebral focal ischemia with reperfusion.【Methods】Thirty SD rats were randomized into 6 groups: sham-operation group,model group,Moschus(1 mg?kg-1?d-1)group,Borneol(3 mg?kg-1?d-1)group,Moschus with Borneol group,nimodipine(12 mg?kg-1?d-1)group.The rat model of cerebral focal ischemia with reperfusion was established with method of putting nylon monofilament in the internal carotid artery.The brain water content was evaluated by detecting the wet weight and dry weight of brain.The detection of cerebral extra-vascular Evans blue(EB)content was used to observe the changes of BBB permeability.【Results】After cerebral ischemia for 2 hours and reperfusion for 24 hours,water content and EB content in the model group increased significantly compared with the pseudo-operation group(P

Expression of the POZ domain of human Zbtb7A and preparation of its polyclonal antibodies / 医学研究生学报

Objective: Proto-oncogene Zbtb7A has been characterized as a molecular switch in the process of cancer initiation and development.Our goal is to obtain the POZ domain of the Zbtb7A protein and prepare its polyclonal antibodies.Methods: We optimized the coding sequence of the POZ domain according to the codon bias of E.coli and synthesized the sequence with two-step PCR,which was then introduced into the pET-26b(+) vector to express the protein.The recombinant protein was analyzed by 15% SDS-PAGE and the corresponding band was cut out from gel.The minced gel slice that contained the POZ domain protein was injected to immunize rabbits.The collected rabbit antiserum was purified using the saturated ammonium sulfate method in combination with protein G antibody purification,and the purified polyclonal antibodies was evaluated by Western blot.Results: The optimized sequence of the POZ domain was correctly obtained and successfully constructed into the pET-26b(+) vector.After induction,an expected protein band about 14 KD was detected on 15% SDS-PAGE,and highly purified polyclonal antibodies were obtained,which were specifically bound to human Zbtb7A.Conclusion: The obtained recombinant protein of the POZ domain and its polyclonal antibodies can be used for further studies of Zbtb7A.

Experimental study of the early period of fracture healing in a rat osteoporotic model prepared by ovariecto-my / 中华骨科杂志

Objective The research was to observe the changes of histomorphology,bone mineral density of callus and the expression and distribution of transforming growth factor beta(TGF-? 1 ),basic fi-broblast growth factor(bFGF)and bone morphogenetic proteins-2(BMP-2)in osteoporotic fracture so as to evaluate the influence of osteoporosis on fracture healing process.Methods An osteoporotic animal model was established by ovariectomy in sixty SD rats,in which the femur was osteotomized by a wire saw as frac-ture healing model,internal fixed with a Kirschner pin three months after ovariectomy.The rats were scari-fied in3days,1,2,4and6weeks after operation,the callus formation was monitored over a6weeks period by histological and immunohistochemistry assessment.Results The status of osteoporosis in ovariectomy group was confirmed by total body bone mineral density measurement 3months after ovariectomy.The aver-age body weight of ovariectomy animals was337.8g compared to286.2g in the control group(P