Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Male ; Middle Aged ; Purpura, Thrombotic Thrombocytopenic ; etiology ; Transplantation, Autologous

Adolescent ; Adult ; Electroacupuncture ; Female ; Garlic ; chemistry ; Humans ; Male ; Middle Aged ; Moxibustion ; Tuberculosis, Gastrointestinal ; therapy ; Young Adult

Objective To explore and establish a kind of dissertation evaluation system which can fully reflects the characteristics of academic medical doctoral training.Methods 41 medical doctoral dissertation evaluation systems and related literatures were collected,to get the current situation of this dissertation evaluation system in our country and find the key points of various evaluation systems.A questionnaire survey was conducted among 350 doctoral supervisors coming from 6 universities so as to define the framework and connotation of evaluation indicators.Using the Delphi expert consultation method,the connotation of evaluation indicators and composition as well as the weight of various indicators was improved.Results The doctoral supervisors who participated in the research considered that building the dissertation evaluation system should follow the principle of science,innovation and fairness,and the evaluation frame-work should have secondary index and evaluation elements.The evaluation contents concluded choosing topic,literature review,basic theory and professional knowledge,paper writing,innovation,research ability and academic morals and so on.And on the basis of this,the paper explored and developed a more scientific and comparative dissertation quality evaluation system with medical specialty.Conclusion The cultivation units should take the educational goal of evaluation target and the requirements of awarding degree into consideration,seeking opinions fully to the first-line supervisors and combing quantitative and qualitative evaluation effectively with scientific and operable principles when specifying evaluation system.

Based on the principles of the in vitro staining technique, hypotonic swelling test, and water test, the Eosin Y-water test method was developed to simultaneously detect the integrity of the sperm head and tail and sperm membrane structure and function. As a widely used method in clinical laboratories in China, the Eosin Y-water test is methodologically characterized by three advantages. Firstly, both the sperm head and tail can be detected at the same time, which allows easy and comprehensive assessment of membrane damage in different parts of sperm. Secondly, distilled water is used instead of the usual formula solution to simplify and standardize the test by eliminating any potential effects on the water molecules through the sperm membrane due to different osmotic pressure or different sugar proportions and electrolyte solutions. Thirdly, the test takes less time and thus can be repeated before and after treatment. This article focuses on the fundamental principles and modification of the Eosin Y-water test and its application in sperm function examination and routine semen analysis for male infertility, assessment of the quality of sperm retrieved by testicular fine needle aspiration, semen cryopreservation program development, and evaluation of sperm membrane integrity after microwave radiation.
Cell Membrane ; China ; Cryopreservation ; Eosine Yellowish-(YS) ; Fluorescent Dyes ; Humans ; Infertility, Male ; diagnosis ; Male ; Osmotic Pressure ; Semen Analysis ; methods ; Sperm Head ; Sperm Motility ; Sperm Tail ; Spermatozoa ; Staining and Labeling ; Water

Objective To determine whether it could enhance gene delivery and tumor transfection in vivo by combination of ultrasound-targeted microbubble destruction(UTMD)with polyethylenimine(PEI)in tumor xenografts.Methods Two different reporter plasmid[lueiferase(pCMV-LUC)and red fluorescent protein(RFP)]were incubated with PEI to prepare cationic compound(PEI/DNA)in various nitrogen:phosphate ratios(N/P ratios,nmol of nitrogen in the PEI/nmol of phosphate in DNA).Formation of PEI/DNA complexes were confirmed by the gel retardation assay.Human cervical carcinoma(Hela)tumors were planted subcutaneously in both flanks of female nude mice.Tumor-bearing mice were administered by tail vein with PBS,plasmid,plasmid and Sono Vue microbubble,PEI/DNA and Sono Vue microbubble.One tumor was exposed to ultrasound irradiation (3 MHz,2 W/cm2,2 min exposure,duty cycle 20%),while the other served as control.The feasibility of targeted delivery and tissue specificity facilitated by UTMD and PEI was investigated.The mice were sacrificed 3 days after ultrasound exposure.Tissue specimens were viewed with microscopy to determine the presence of RFP expression.The efficiencies of luciferase transgene expression were determined.Histology analysis was detected.Results Electrophoresis experiment revealed that PEI was mixed with plasmid to condense DNA efficiently.The application of UTMD significantly increases the tissue transfection in vivo compared to plasmid alone.RFP expression was present in all sections of tumors that received ultrasound exposure but not in control tumors.Results of luciferase activity showed that the expression of luciferase was to be 14 times greater in ultrasound-exposed tumors(P＜0.01).More importantly,the increase in transgene expresgion was related to UTMD with the presence of PEI dramatically.At least 10-fold increase of luciferase gene transfer was obtained in irradiated tumors compared to non-irradiated controls(P＜0.01),111-fold increase compared to UTMD alone(P＜0.01).There was not significantly gene expression in other organs or tissues regardless of US exposure(P＞0.05).No tissue damage was seen histologically.Conclusions The combination of UTMD with PEI can enhance targeted delivery and expression of reporter gene to tumors at intravenous administration.It is a promising new and safe method for gene delivery in vivo.

Objective: To optimize the formula and preparation technology of cantharidin chitosan bioadhesive microspheres.Methods: The microspheres were prepared by a spray drying method.A single-factor experiment was used to study the effect of chitosan with different molecular weight on the gastric mucosa adhesion rate were also studied, and the effects of different drug loading ratio, concentration of chitosan acetic acid solution and speed of peristaltic pump on the loading rate were also studied.The cantharidin entrapment efficiency as the studying index, a response surface method was used to further optimize the formula.Results: The best formula and preparation process of cantharidin chitosan bioadhesive microspheres were as follows: the concentration ratio of antharidin to chitosan was 19.83% , the concentration of chitosan acetic acid solution was 0.77% , and the peristaltic pump flow rate was 9.225 ml·min-1.With the best formula, the entrapment efficiency of cantharidin chitosan bioadhesive microspheres was 90.14%.Conclusion: The microsphere preparation by the spray drying method is stable and repeatable.Cantharidin chitosan bioadhesive microspheres have high entrapment efficiency and promising biological adhesion.

Objective To investigate the diagnostic value of contrast-enhanced ultrasound (CEUS), and compare it with contrast-enhanced CT(CECT). Methods Eighty-one focal liver lesions were evaluated with CEUS and CECT, diagnostic value of two modalities were calculated, vaseularity manifestations of two modalities in arterial phase were compared. Results The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of CEUS were 96.6 %, 86.9 %, 94.9 %, 90.9 %, 93. 80%, respectively, and the diagnostic value of CECT were 98.3%, 82. 6%, 93.4%, 95.0%, 93.8%, respectively. Among all the metastasis, hypervascular manifestation lesions in arterial phase of CEUS and CECT were 72% (18/25) and 32% (8/25)( P <0.05). Conclusions The diagnostic value of CEUS and CECT in focal liver lesions are comparably satisfying,and there is no significant difference between them, the combination use of two modalities would improve the diagnosis performance. CEUS can show more hypervaseular metastasis than CECT in arterial phase.

Objective To explore the optimal dose-effect relationship of gene transfer according to the expression of pEGFP in tumor cells under different parameters mediated by microbubble contrast agent and ultrasound in vivo.Methods C57BL/6J mice were inoculated subcutaneously in the middle of the right flank with hepal-6 tumor cells.The mixture of SonoVue and enhanced green fluorescent protein(EGFP)plasmid was injected into the tail vein of each mouse, groups were randomly established according to different output intensity( 1 W/cm2, 2 W/cm2, 3 W/cm2 ), exposure time ( 1 min, 5 min, 10 min) and volume of SonoVue(30 μl,60 μl,90 μl), the expression of EGFP in tumor cells under different parameter was examined by flow cytometry and fluorescence microscopy.Results The expression of pEGFP in tumor cells was significantly higher with the increase of exposure time,output intensity and volume of SonoVue (1 min vs 5 min, P＜0.05; 1 W/cm2 vs 2 W/cm2, P＜0.05 ; 30/μl vs 60 μl, P＜0.05).However, the transfection efficiency of EGFP can not be enhanced continuously with the increasing exposure time,output intensity and volume of SonoVue (5 min vs 10 min, P＞0.05; 2 W/cm2 vs 3 W/cm2, P＞0.05; 60 μl vs 90 μl, P＞0.05).The highest transfection effieieney was achieved under intensities was 2 W/cm2 [(21.02±1.45)%]with 5 min[(23.22±1.91)%] exposed to ultrasound, and 60 /μl SonoVue [ (21.02±1.45) % ].Conclusions Gene transfeetion efficiency changded with the difference parameters.2 W/cm2,5 min and 60 μl SonoVue led to the optimum dose-effect relationship of gene transfer in vivo .

Objective To probe into the clinical value of left atrial(LA) volme tracking method (LAVT) on the assessment of left atrial function in patients with hypertension. Methods Thirty-one patients with hypertension and 31 control subjects were involved. Images of apical two-chamber and four-chamber views were obtained by two-dimensional echocardiography. LA maximal volume (LAVmax), LA minimal volume (LAVmin), LA presystolic volume (LAVpre), systolic LA filling rate (dv/dtS), early diastolic LA emptying rate (dv/dtE) and late diastolic LA emptying rate (dv/dtA) were derived using LAVT. LA passive emptying volume(LAVp), LA passive emptying fraction(LAVpEF), LA active emptying volume(LAVa), LA active emptying fraction (LAVaEF), LA total emptying volume (LAVt), LA total emptying fraction(LAVtEF) were calculated. All volume measurements were corrected to body surface area (LAVI). Results There were significant differences in the LAVImax, LAVImin, LAVIpre, LAVpEF, LAVIa, LAVIt,dv/dtS, dv/dtA between hypertensive patients and control subjects(P <0.05~0.001), but no significant differences were found in the LAVp, LAVaEF, LAVtEF,dv/dtE between the two groups. The LAVImax has shown significant positive correlation with LAVIp, LAVIa and LAVIt(r = 0.588~0.812, P <0.001). LAVaEF has shown positive correlation with the LAVtEF(r = 0.833, P<0.001), whereas LAVpEF has not(r = 0.420, P <0.01). There was a significant positive correlation with LAVImax and LAVIpre(r = 0.908, P <0.001), LAVIpre and LAVIa(r = 0.689, P <0.001) ,dv/dtE and LAVIp(r = 0.690, P <0.001),dv/dtA and LAVIa(r = 0.600, P <0.001). Conclusions In hypertensive patients,left atrial reservoir function and booster pump function increase, while left atrial conduit function decreases. LAVT has a potential ability to evaluate left atrial function.

Objective To study the transfeetion efficiency and safety of liposome microbubble(LM)on red fluorescent protein(RFP)in vitro and in vivo under ultrasound mediated gene transfection(USMGT)conditions.Methods Plasmids containing RFP were added to cultured Hela cells followed by ultrasound (US)exposure with LM.Different concentration of LM,US intensity and exposure time were optimized.Transfection efficiency was evaluated by fluorescent microscopy and FACS.Cell viability was verified by propidium iodide assay.In transplanted tumors in vivo study,LM and plasmid(P)were injected into the nude mice followed by US exposure(P+LM+US group).Nude mice undergoing plasmid injection alone(P group),plasmid injection and US exposure(P+US group)and plasmid and LM injection(P+LM group)were used as controls.Frozen section and histological examination were conducted and RFP expression was evaluated.Results LM and US exposure significantly increased transfeetion efficiency in cultured Hela cells (P＜ 0.01).Transfection efficiency was the most prominent under the condition of US intensity of 1.0 W/cm2 with 6%LM,duration 3 min.No apparent cell damage was found in the all groups.In transplanted tumors,strong RFP was seen in P+LM+US group.It was significantly higher than in any other groups(P＜0.0 1).No tissue damage was seen histologically.Conclusions LM could enhance USMGT effectively without causing any apparently adverse effect in vitro and in vivo.This method would be a novel,effective,safe non-viral gene transfection method and provide an alternative to current clinical gene therapy.