Objective To develop a novel approach of transfecting the exogenous gene safely and effectively mediated by receptor according to the cavitation effect of ultrasound-mediated microbubble destruction. Methods C-myc antisense oligonucleotide(ASODN) and galactosylation polylysine(G-PLL) which was fluoresce in-labeled specificity ligand and SonoVue microbbule were coupled by electrostatic interaction. SMMC-7721 cell with c-myc ASODN were transfected in vitro by ultrasound-mediated microbubble destruction to investigate the effect of c-myc ASODN on it. The c-myc mRNA gene expression of liver cancer SMMC-7721 cells was detected. Results Cultured with 10% SonoVue microbubble and under ultrasound at 1.5 MHz,1.0 W/cm2, 10% duty cycle,60 s respectively,the SMMC-7721 cells had not been changed significantly. Reverse transcription polymerase chain reaction(RT-PCR) assay showed that c-myc expression was decreased in transfected cells(P<0.01). Conclusions Receptor-mediated targeted microbubble destruction by ultrasound can enhance the exogenous gene transfection and expression. It is a promising method in gene therapy of liver cancer.

A co-words network for high frequency words was established using the title information of papers pub-lished in 22 medical journals from 2005 to 2013.The evolution of medical scientific research topics was analyzed with the visual analysis software NEViewer, which showed that medical research in China was focused on basic and application studies with the prevention and treatment of frequently encountered diseases and the associated technical methods as the two main directions, and that medical scientific research topics would remain their stable status in a rather long period.The hot medical topics were explained.

Background and purpose:Thermal ablation (radiofrequency ablation, RFA/microwave ablation, MWA) is the most commonly used minimally invasive technique for the treatment of liver cancer. Real-time shear wave elastography (SWE) is a new type of ultrasonic imaging technology, which was used in our study to monitor thermal ablation of liver cancer. This study aimed to investigate the stiffness change of liver cancer and that of surrounding liver parenchyma before and after thermal ablation, and to evaluate the application of SWE for monitoring thermal ablation for liver cancer.Methods:From Oct. 2014 to Apr. 2015, a total number of 36 patients, with 39 lesions, were treated with RFA or MWA and got complete response. SWE examination was performed before and after ablation. The SWE-mean, SWE-min, SWE-max, SWE-SD of lesions and the surrounding liver parenchyma were measured. Statistical analysis was made to compare the stiffness changes of liver cancer with those of the surrounding liver parenchyma before and after thermal ablation, and to determine whether there were differences between two different ablation modes.Results:Before and after ablation, the SWE-mean of lesions was (30.09±11.67) kPavs (52.11±17.56) kPa,SWE-min was (10.46±8.22) kPavs (20.57±11.42) kPa, SWE-max was (51.50±20.84) kPavs (88.54±27.75) kPa, SWE-SD was (10.63±4.30) kPavs (16.89±7.72) kPa; There were statistically signiifcant differences (P<0.05). Before and after ablation, the SWE-mean of surrounding liver parenchyma was (8.84±2.82) kPavs (8.91±2.78) kPa, SWE-min was (4.77±1.95) kPavs (4.69±1.90) kPa, SWE-max was (13.82±3.79) kPavs (14.34±3.97) kPa, SWE-SD was (3.24±1.32) kPavs (3.37±1.29) kPa; There were no statistically signiifcant differences (P>0.05). After ablation, the SWE-mean of RFA and MWA was (45.55±10.91) kPavs (60.59±20.99) kPa, SWE-min was (18.95±8.86) kPavs (25.93±10.93) kPa, SWE-max was (76.58±15.51) kPavs (104.01±32.59) kPa, SWE-SD was (13.82±3.52) kPavs (20.85±9.77) kPa; There were statistically signiifcant differences (P<0.05).Conclusion:SWE can quantitively analyze the stiffness of lesions. The ablation zone became stiffer after RFA or MWA, and the ablation zone of MWA was stiffer than that of RFA. Two kinds of ablation methods did not signiifcantly affect the stiffness of liver parenchyma around the lesion. SWE could potentially be used to monitor thermal ablation of liver cancer.

Objective To explore the features of high-frequency ultrasonography and contrast-enhanced ultrasonography (CEUS) of papillary thyroid microcarcinoma (PTMC).Methods The CEUS data and ultrasound data of 147 PTMCS which were reconfirmed by pathology were analyzed retrospectively,and the CEUS and ultrasonic characteristics of them were summarized.Results Among 147 nodules,144 (97.9％) nodules were hypoechoic,and 3 nodules were isoechoic.Vague edge was found in 136(92.5％) PTMCs,and 126(85.7％) PTMCs were irregular in shape.Totally 92(62.6％) PTMCs were A/T ＞ 1,microcalcifications were found in 81 (55.1％) PTMCs.Besides,26(74.2％) PTMCs were found microcalcification in 35 PTMCs combined with Hashimoto's thyroiditis (HT),while 55 (49.1％) PTMCs were found microcalcification in 112 PTMCs combined with HT.There were significant differences between them (P ＜ 0.05).The blood distribution of 129 (87.8％) nodules was type Ⅱ.The contrast-enhanced pattern of 147 (100.0％) PTMCs showed in-homogeneous enhancement in 144 (97.9％) nodules,hypoenhancement in 136(92.5％) nodules,and all the nodules without amicula.Conclusions The typical PTMCs are hypoechoic,irregular shapeand vague edge,usually were found as A/T ＞ 1,microcalcification,and type Ⅱ blood distribution.With the method of contrast-enhanced ultrasonography,these nodules usually without amicula showed inhomogeneous and hypoenhancement.The incidence of microcalcification is more common when patients with Hashimoto's disease coexisting PTMC.

The effects of several factors on the chlamydospores production of Trichoderma aureoviride T-33 during the fermentation were researched.Based on the results above, the orthogonal test was made to screen out the best prescription.The results showed that, the best single-factor conditions for the chlamydospores production of T.aureoviride T-33 were, liquid culture of oat powder, 30℃, pH4.0, 120r/min, 24 hours oscillate incubating as well as liquid culture volume of 80mL/bottle when the 250mL size triangle bottle was used.The result of orthogonal test showed that, the best prescription for temperature, pH and oscillating speed was 30℃, pH4.0 and 140r/min.3.37?10~ 7 spore/mL chlamydospores were obtained at this combined condition.

Objective To observe the curative effect in patients with systemic lupus erythematosus(SLE) complicated by moderate or severe thrombocytopenia with a blood platelet count(BPC) of under 50 × 109/L and analyze its related factors. Methods We retrospectively analyzed the clinical data on 109 SLE patients with mod-erate or severe thrombocytopenia. Results Of the 109 patients,82(75.2%)had complete response(CR),15 (13.8%)had partial response(PR),and 12(11.0%)had no response(NR),respectively. As compared with the CR+PR group,the NR group had a higher incidence rate of decreased bone marrow megakaryocyte(P < 0.05). However,there were no significant differences between the two groups in SLEDAI scores,rates of positive PAIg and ACA,and levels of C3 and C4(P>0.05 for all comparisons). The total effectiveness rate did not differ signifi-cantly between MP pulse therapy and high-dose corticosteroid therapy. Conclusions A decrease in bone marrow megakaryocytes can be an adverse factorfor affecting the efficacy in patients with SLE complicated by moderate or severe thrombocytopenia.

Aim To investigate the effect of the GABAA receptor on the analgesia,hypnosis and amnesia induced by GABA in mice.Methods GABA was intracerebroventricularly(icv) injected and securinine,a GABAA receptor antagonist,was intravenously(iv) injected after the GABA intracerebroventricularly injection.Then four techniques including hot-plate test,acetic acid-induced writhing test,awaken test as well as step-through test & step-down test were employed to evaluate the effects on the pain index in hot-plate test(HPPI),the writhing times,the sleeping time(ST) and the latency & the number of errors.Results GABA increased the HPPI and ST,but decreased writhing times and the latency & increased the number of errors in conscious mice;the above effects are dose-dependent.Securinine could antagonize the effects stimulated by GABA.Conclusion GABA lead to analgesia,hypnosis and amnesia.GABAA receptor might be the important target for these effects.

ObjectiveTo prepare high specific monoclonal antibodies(mAbs) against BP26 of Brucella(B.)melitensis.Methods A recombinant plasmid pET-28a-BP26 was constructed and transformed into competent Escherichia coli BL21 (DE3),and then the bacteria were induced by 1 mmol/L isopropylthio-β-D-galactoside (IPTG).After induction,the recombinant BP26 protein (rBP26) was purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PGAE) and nickel ion affinity chromatography(Ni-NTA).Mice were inoculated with rBP26 antigens for three times at 2-week intervals.The first subcutaneous injection contained 100 μg rBP26 with 0.1 ml complete Freund adjuvant.The second subcutaneous injection was 50 μg rBP26 with 0.1 ml incomplete Freund adjuvant.The antibody titers to rBP26 were determined 2 weeks after each reimmunization.Three days before cell fusion,the mice with the highest titer were intraperitoneally injected with 50 μg rBP26 in 0.1 ml PBS.Pre- and post-immunization sera were collected and used as negative or positive controls for screening mAbs.Mice with the highest titer were sacrificed and spleen cells were isolated.The spleen cells of rBP26 immunized mice were fused with SP2/0 myeloma cells in a ratio of 5 ∶ 1 by polyethylene glycol(PEG) 1450.Antibody-producing hybridomas were primarily screened by an indirect enzyme-linked immunosorbnent assay(ELISA) with rBP26.Reactive hybridomas were subcloned for 3 times,then the strains of hybridoma cells secreting antibodies against BP26 were obtained.Supernatant of cloned hybridoma cultures was collected for mAb analyses.These mAbs were named by the hybridoma clone number and tested their reactivity to membrane proteins extracted(NMP) from B.melitensis vaccine strain(M5-90) by Western blotting and Dot-ELISA.mAbs isotyping and kappa(κ) or lambda(λ) light chain was identified by Mouse Monoclonal Antibody Isotyping Kit.Results A total of two mAbs reactive to rBP26 of B.melitensis were selected from antibody screening hybridomas by indirect-ELISA.The two mAbs were named 3C3 and 5A5,and identified as IgG1 (κ) and IgG2(κ),respectively.They could react with NMP from M5-90.Conclusions Results of identification show that two mAbs against rBP26 can be produced.The two mAbs can recognize natural BP26 protein,giving the experimental materials for further research on identification of its epitopes.

A BBB co-culture cell model consisting of rat brain microvascular endothelial cells (BMEC) and astrocytes (AS) was established to study the effect of Angelica dahurica coumarins on the transport behavior of puerarin across blood-brain barrier (BBB) in vitro and in vivo. The barrier function of this model was evaluated by measuring the transendothelial resistance, phenol red permeability and BBB related protein expression. The permeability assay and western blot methods were performed to study the effects of Angelica dahurica coumarins on the BBB permeability and the expression of BBB related protein. The animal experiment protocols in this study were approved by the Animal Ethics Committee of Xi'an Jiaotong University (Animal Ethics No.: 2021-1329). The results showed that the established BMEC/AS co-culture model could be used to evaluate drug transport across BBB in vitro. After combined with Angelica dahurica coumarins, the transport capacity of puerarin was significantly increased in vitro and in vivo. Additionally, Angelica dahurica coumarins enhanced BBB permeability and inhibited the protein expression of P-glycoprotein (P-gp), zonula occludens-1 (ZO-1) and occludin. Angelica dahurica coumarins might increase BBB permeability by inhibiting the expression of P-gp and tight junction protein, thereby increasing the content of puerarin in brain tissue.

ObjectiveTo study the effect of enriched rehabilitative training on the functional recovery and neuronal dentritic growth following cerebral ischemia/reperfusion.Methods32 male Wistar rats,weighting 180～200 g,were randomly divided into a ischemic group(n=16) and a sham-operation group(n=16) after beforehand trainings.Rats were subjected to 2 h of right middle cerebral artery occlusion before reperfusion.After surgery,the ischemic group were randomly divided into a ischemia + enrichment(IE) group and a ischemia + standard housing(IS) group;the sham-operation group were randomly divided into a sham + enrichment(SE) group and a sham + standard housing(SS) group.After 24 h reperfusion,IE and SE groups were housed in enriched cages,and given enriched rehabilitative training according to the scheme.At the same time,IS and SS groups were housed in standard cages without any training.The functions of 4 groups were evaluated at 24 h,1 week,2 weeks,3 weeks and 4 week after operation.Dentritic growth of layer V pyramidal cells of the undamaged forelimb motor cortex was examined using Golgi-Cox procedure.ResultsIE group showed better function than IS group in all behavioral test.There was no significant difference in limb-placement test at 3 weeks(P>0.05) and in footfault test at 4 weeks(P>0.05) after operation between IE and SE group.The mean of basilar dentrite branching points in IE group was significantly greater than that of other groups(P<0.01).ConclusionEnriched rehabilitative training can promote functional recovery and enhance neural plasticity after cerebral ischemia/ reperfusion in rats.