ObjectiveThe aim was to construct the recombinant plasmid of pET-28a-G-protein pathway suppressor 2 (GPS2) GPS2,express GPS2 protein in E.coli,and obtain specific polyclonal antiserum of GPS2.MethodsGPS2 gene was obtained and the amplified fragment was then cloned into E.coli expression vector pET-28a to construct recombinant plasmid.The recombinant plasmid was transformed into E.coli expression strain BL21(DE3).IPTG induces the expression protein GPS2 protein,and the induction conditions were optimized.The induced product was purified by Ni2+ affinity chromatography,and the purified product was dialyzed with buffer for refolding.The purified protein can be used as antigen,injected to immunize male New Zealand white rabbit to get polyclonal antiserum.The titer and specificity of the rabbit antiserum were detected by ELISA and Western Blotting.ResultsThe E.coli expression vector pET-28a-GPS2 was constructed successfully and the recombinant protein was efficiently expressed and purified.The purified protein was used to immunize male New Zealand white rabbit to get polyclonal antiserum and the ELISA and Western Blot results showed that the high titer of specific polyclonal antiserum.ConclusionGSP2 could be highly expressed in E.coli.Antiserum of GPS2 protein can be obtained by the purified recombinant to analyze its function.

Objective To discuss the ultrasonographic characteristics of schistosomal appendicitis lesions. Methods Among the patients with schistosomal hepatopathy who were discovered by Color Doppler ultrasound in Huzhou Central Hospital from January 2012 to December 2015,50 cases with clear history of schistosomiasis and treatment were chosen as a schistosom?al hepatopathy group,meanwhile,50 normal people,who came from non?endemic areas,without schistosomal hepatopathy and schistosomiasis history were chosen as a control group. The two groups were examined by ultrasound scan of the appendix ,and the data of the largest diameter of the appendix and the thickness of the appendix wall were collected,and the sonographic char?acteristics of their appendixes,such as whether the echo of the appendix wall was even or not,were observed. Results The minimum internal diameter of the appendix cavity and the thickness of the appendix wall of the schistosomal hepatopathy group were(2.090 ± 0.790)mm and(1.332 ± 0.313)mm ,respectively,the former was significantly narrower than that of the control group,while the latter was significantly thicker than that of the control group(t=2.647,-4.526,respectively,both P<0.05). The proportions of those with inhomogeneous echo,indistinctness structure,uneven thickening of the appendix wall,as well as having intestinal contents in the appendix cavity in the schistosomal hepatopathy group were higher than those in the control group(χ2=12.000,18.537,24.008,4.244,respectively,all P<0.05). Conclusions Schistosomal appendicitis lesions have obvious ultrasonographic characteristics under ultrasound. Ultrasound can play an important role in judging whether the appen?dix of schistosomiasis patients is involved and discovering the lesion of appendix early.

Objective To prepare a redox-responsive doxorubicin-loaded nanoparticle,and to study its in vitro realease behavior and targeting effect on heptoma cells.Methods Cystamine was grafted on the side chains of hyaluronic acid with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide catalyst,and then β-cyclodextrin (β-CD) was conjugated on the amine groups of the cystamine by Schiff's base reaction to prepare β-CD modified hyaluronic acid (HACD).The HACD/DOX nanoparticles were prepared by encapsulating DOX into HACD using dialysis method.The drug loading,encapsulation efficiency,particle size and distribution,zeta potential and other physical and chemical properties,as well as in vitro drug release behavior of the HACD/DOX nanoparticles were characterized.The cytotoxicity of HACD/DOX nanoparticles to HepG2 cells was studied by cell counting kit-8 (CCK-8) method.The targeting effect of HACD/DOX nanoparticles on HepG2 cells was studied using flow cytometry and confocal laser scanning microscopy (CLSM).Results HACD were successfully synthesized,which could carry DOX to form uniform homogeneous nanoparticles.The drug loading of DOX in the nanoparticles was (16.1±0.2)％ and the encapsulation efficiency was (64.2±0.9)％.The transmission electron microscope images indicated that the shape of the HACD/DOX nanoparticles was homogeneous sphere.The results of granularity analysis showed that the average size of the HACD/DOX nanoparticles was (203.1 ±2.5) nm with a narrow size distribution (PDI =0.202).The zeta potential of the HACD/DOX nanoparticles was (-29.1±0.8) mV.The in vitro release behavior of the nanoparticles exhibited obvious redox-sensitivity.The results of in vitro cytotoxicity showed that the blank carrier material HACD had no obvious toxicity to hepatoma cells,and the HACD/DOX nanoparticles could effectively kill hepatoma cells with the 0.38 μg/ml half maximal inhibitory concentration (IC50) value at 48 h.Flow cytometry and CLSM results demonstrated that the HACD/DOX nanoparticles could target hepatoma cells through the mediating effect of hyaluronic acid.Conclusions The prepared HACD/DOX nanoparticles have suitable particle size,high drug loading and encapsulation efficiency,and can release DOX under the stimulation of reducing agent.These nanoparticles have obvious targeting effect on hepatoma cells,which is expected to be applied as the drug delivery system of hepatocellular carcinoma (HCC) therapy.

Objective To evaluate efficacy ,stability and safety after implantation of toric implantable collamer lens (TICL) to correct moderate to high myopic astigmatism .Methods This study evaluated 125 eyes of 71 patients with moderate to high myopic astigmatism who accepted TICL implantation .LogMAR uncorrected(UCVA) and best corrected(BCVA) visual acuity ,intraocular pressure ,cycloplegic refraction and manifest refraction examination were assessed preoperatively on 3 ,6 ,12 months postoperatively . The rotation of TICL axis were measured on 3 ,6 ,12 months postoperatively .Results 12 months postoperatively ,the logarithm of the minimum angle of resolution(LogMAR) UCVA were 0 .064 ± 0 .157 which were remarkably better than preoperative BCVA (P<0 .01) .119 eyes(95 .2% ) had postoperative UCVA better than or equal to preoperative BCVA .The manifest spherical refrac‐tion(absolute value) was (0 .36 ± 0 .41)D .105 (84 .0% ) eyes were within ± 0 .5 D .The mean manifest refractive cylinder was (-0 .63 ± 0 .61)D .109(87 .2% ) eyes had ≤ -1 .00 D .The mean rotation on 12 months postoperatively was (4 .76 ± 6 .14)° .The rotation of 101(80 .8% ) eyes were within 5 degrees .No vision threatening complications occurred during the observation period . Conclusion Implantation of TICL is safe and stable in the treatment of moderate to high myopic astigmatism .TICL is an ideal sur‐gical option to treat moderate to high myopic astigmatism .

Objective To compare the skills level before and after pediatric advanced life support course and analyze the effect of the training. Methods The pediatric advanced life support was used as the textbook. The skills were got through attending theory classes, watching demonstrations and taking part in the simulator training. The questionnaires were filled strictly and the data was analysed. Results The test scores were increased after the training (P＜0. 01). There were only 8.7% of the trainees had used the rescue equipments and 61.3% had never seen the rescue equipments before training. More than 80% of the trainees were satisfied with the training about the utility and novelty. Conclusion pediatric advanced life support course can successfully deliver a large number of healthcare providers with international unique pediatric emergency treatment skills ,and raise the participants abilities of rescuing critical children.

Objective To explore the feasibility and value of dynamic contrast-enhanced (DCE-MRI) parameters in assessing early effects of anti-angiogenesis medicine in targeted therapy for tumors.Methods Twenty BALB/C-nu nude mice were injected subcutaneously with human colon cancer cells HT-29 to the right hind leg.The nude mice were evenly divided into the experimental group and control group with 10 mice in each group.The mice of experimental group were intraperitoneally injected with bevacizumab,and the control group were injected with the same volume of saline.DCE-MRI was performed before medication and one hour,24 h and 48 h after medication.The Ktrans,Kep,Ve and initial area under enhancement curve (iAUC) of DCE-MRI were analyzed.The animals were sacrificed 48 hours after medication.Microvessel density (MVD) of the tumors was detected by immunohistochemistry.One way analysis of variance was performed to analyze parameters of DCE-MRI.The Pearson correlation coefficient was used to analyze the correlation between parameters of DCE-MRI and MVD.Results Under DCE-MRI,the edge of subcutaneous colon cancer xenografts was obviously gradually enhanced,pseudo color indicated high perfusion,the strength degree of the central region was low and which meant low perfusion.The differences in Kep of different time point of experimental group were statistically significant (F=3.752,P=0.016) ; there as no significant difference in other parameters of DCE MRI (all P＞0.05).There was no significant difference in Ktrans and Kep before medication and one hour after medication (all P＞0.05).There were significant difference in Ktrans and Kep 24 hour and 48 hour after medication between experimental group (24 hour∶ (0.095 ± 0.039) min-1 and (0.297 ± 0.141) min-1,48 hour∶ (0.090±0.033) min 1 and (0.314±0.148) min-1) and control group (24 hour∶ (0.150±0.074) nin-1 and (0.494±0.126) min-1,48 hour∶ (0.171±0.045) min-1 and (0.441± 0.092) min-1) (F24h =4.824 and 11.386,F48h =22.605 and 5.455,all P＜0.05).There was no significant difference in Ve and iAUC between two groups at different time points (all P＜0.05).MVD of experimental group was lower than that of control group.Ktrans and Kep were positively correlated with MVD (r=0.745 and 0.400,both P＜0.05).Conclusion Ktrans and Kep parameters of DCE-MRI may be used in monitoring the earlier effects of anti-angiogenesis medicine in targeted therapy for colon cancer.

Objective To explore the value of spectral imaging technique in dual-energy CT in differential diagnosis of the metastatic lymph nodes in thyroid carcinoma,squamous cell carcinoma metastatic lymph nodes and lymphoma in the neck.Methods In 30 patients with pathologically confirmed with a total of 79 cervical lymph nodes enlargement which were using dual energy scan .Then observed the change trend of the spectrum curve and comparison the three kinds of lymph node energy spectrum curve’s slope.Results In the 79 lymph nodes,the metastatic lymph nodes in thyroid carcinoma were twenty-three,squamous cell carcinoma metastatic lymph nodes were twenty-four and lymphoma were thirty-two.From 60 to 180 keV,with the increase of keV values,the three kinds of malignant lymph nodes of the corresponding CT value decreasing and the higher the keV value,the CT value decrease magnitude was small,and the spectrum curve was〞drop type〞.The slope spectrum curve of the metastatic lymph nodes of thyroid carcinoma in arterial phase and parenchymal phase were maximum,which were 1.23±0.41 and 0.85±0.33,respectively.The slope spectrum curve of lymphoma in arterial phase and parenchymal phase were least,whcih were 0.40±0.16 and 0.47 ±0.09.The slope spectrum curve of the squamous cell carcinoma metastatic lymph nodes in arterial phase and parenchymal phase were 0.88±0.10 and 0.62±0.28.The energy spectrum curve slope of the three kinds of malignant lymph nodes have statistical significance.Conclusion The energy spectrum curve slope of arterial phase and parenchymal phase has some significance lymph node metastasis of in thyroid carcinoma,the metastatic lymph nodes and lymphoma in the neck.

BACKGROUND:Human hypoxia-inducible factor-1 alpha can regulate the expression of osteogenic and angiogenic genes, and promote osteogenic activity. OBJECTIVE:To observe the expression of osteogenic genes in rat bone marrow mesenchymal stem cells carrying human hypoxia-inducible factor-1 alpha slow virus infection. METHODS:Hypoxia-inducible factor-1 alpha was obtained from Hela cells using RT-PCR. Lentivirus expression vector plasmid carrying hypoxia-inducible factor-1 alpha (Lenti-HIF-1α-eGFP) was constructed. 293Ta cells with LentiPac HIV mixed packaging plasmid was packaged, and then lentivirus was obtained. Rat bone marrow mesenchymal stem cells were isolated and cultured using direct whole bone marrow adherent method. Bone marrow mesenchymal stem cells were identified using flow cytometry. Bone marrow mesenchymal stem cells were infected with slow virus for 1, 4, 7 and 14 days. Bone morphogenetic protein-2, osteocalcin, osteopontin and alkaline phosphatase expression levels were detected in bone marrow mesenchymal stem cells using real-time fluorescent quantitative PCR. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cells were effectively infected with Lenti-HIF-1α-eGFP. Real-time fluorescent quantitative PCR results revealed that bone morphogenetic protein-2, osteocalcin, osteopontin and alkaline phosphatase began to obviously overexpress from 4 days after infection with Lenti-HIF-1α-eGFP until 14 days. Results suggested that hypoxia-inducible factor-1 alpha could elevate the osteogenic activity of bone marrow mesenchymal stem cells.

Objective:To investigate the correlation between time within the glucose target range(TIR) and hyperuricemia(HUA) in patients with type 2 diabetes mellitus(T2DM).Methods:A total of 215 patients with T2DM in the First Affiliated Hospital of Bengbu Medical College from June 2021 to May 2022 were selected and divided into HUA group and non-HUA group according to serum uric acid level. The clinical characteristics and biochemical indicators of the patients were collected. The association of 72 h glucose monitoring system(FGMS) related indicators TIR, mean blood glucose fluctuation range(MAGE), blood glucose variability(CV), blood glucose standard deviation(SDBG), and mean blood glucose(MBG) with serum uric acid level was analyzed. The influencing factors of T2DM combined with HUA were analyzed with binary logistic regression, and receiver operating characteristic(ROC) curve was drawn to evaluate their predictive values.Results:TIR of HUA group was significantly decreased compared with non-HUA group, while HbA 1C, MAGE, CV, SDBG, and MBG were increased( P<0.001). Spearman correlation analysis showed that serum uric acid levels were negatively correlated with TIR, but positively correlated with MAGE, CV, SDBG, and MBG( P<0.001). After dichotomous logistic regression analysis, TIR was found to be an independent protective factor for T2DM with HUA. The ROC curve results showed that the area under the curve(AUC) of TIR for predicting HUA in T2DM was 0.856(95% CI 0.803-0.909, P<0.001), with the best cut-off value being 64.5%, the sensitivity being 76.8%, and the specificity being 90.3%. Conclusion:TIR in patients with T2DM combined with HUA was significantly decreased. TIR is an independent protective factor for T2DM combined with HUA, and TIR shows a certain predictive value for T2DM combined with HUA.

Objective:To investigate the efficacy and safety of accelerated epithelium-off corneal collagen cross-linking (CXL) in the treatment of corneal ectasia after keratorefractive surgery.Methods:An observational case series study was performed.Twelve patients (22 eyes) diagnosed with corneal ectasia after keratorefractive surgery in the First Affiliated Hospital of Army Medical University were enrolled from January 2016 to December 2018.All the patients received accelerated epithelium-off CXL and were followed up for 12 months.Before and 1 week, 1, 3, 6, and 12 months after the operation, the uncorrected visual acuity (UCVA) and best corrected visual acuity (BCVA) converted to the logarithm of the minimum angle of resolution (LogMAR) unit were measured.The sphericity, cylindricity, and spherical equivalent were examined by Topcon auto refractor.The maximum keratometry (Kmax) of the front surface, mean keratometry (Km) of the front surface, Km of the back surface, symmetry index of front surface (SIf), symmetry index of back surface (SIb), thinnest corneal thickness (TCT), total aberrations, total high-order aberrations, coma aberration, trefoil aberration and spherical aberration were detected by the Sirius analyzer.The depth of corneal demarcation lines was determined by optical coherence tomography.The intraocular pressure was measured by the non-contact tonometry.The corneal endothelial cell density was assayed by the endothelial cell densitometry.The inflammatory reaction and haze were observed with a slit lamp at different time points after surgery.This study adhered to the Declaration of Helsinki.The study protocol was approved by the First Affiliated Hospital of Army Medical University (No.KY2020063). Written informed consent was obtained from each patient before entering the cohort.Results:Among the 22 eyes of 12 cases, 3 eyes of 2 cases (13.64%) underwent small incision lenticule extraction, and 19 eyes of 10 cases (86.36%) underwent excimer laser in situ keratomileusis.The UCVA (LogMAR), BCVA (LogMAR), cylindricity and spherical equivalent before the operation were 0.61±0.42, 0.24±0.23, (-2.83±2.39)D, (-3.60±2.66)D, which were significantly worse than 0.45±0.31, 0.12±0.15, (-2.11±1.67)D, (-3.12±2.31)D at 12 months after the operation ( t=4.054, 4.956, -3.728, -2.742; all at P<0.05). The front surface Kmax, front surface Km and SIf at 12 months after the operation were (48.37±5.80), (41.49±3.04), (5.36±4.07)D, which were significantly lower than (49.61±5.97), (41.66±2.97), (5.85±4.18)D before the operation ( t=5.949, 2.278, 2.719; all at P<0.05). There was no significant difference in sphericity, Km of the back surface, SIb, TCT, total aberrations, total high-order aberrations, coma aberration, trefoil aberration, spherical aberration, intraocular pressure and endothelial cell density between before and 12 months after the operation (all at P>0.05). Grade 0.5-2 haze occurred in 8 eyes of 4 patients one month postoperatively.After administration of prednisolone acetate eye drops, haze decreased or disappeared 3 months postoperatively, with UCVA and BCVA unchanged.A corneal demarcation line with a depth of (285.40±51.61)μm was found in 11 eyes of 6 cases at 1 month after operation. Conclusions:Accelerated epithelium-off CXL can significantly improve visual acuity, reduce corneal astigmatism and corneal curvature, as well as effectively prevent the progress of corneal ectasia.