Abstract:Objective：To develop a novel method for treating complete heart block by transplanting autologous sinoatrial node（SAN） cells to the right ventricle adjacent to heart apex. Methods： Twelve young pigs were evenly randomized into transplantation group and control group. The sinoatrial nodes of all pigs were harvested and SAN cells were isolated in vitro. Pigs in transplantation group were injected with a cell suspension and those in control group were injected with culture medium into autogeneic right ventricular wall adjacent to heart apex after the temporary myocardial pacing wires were fixed. Two weeks later, all pigs underwent electrophysiology mapping and transcatheter ablation of His bundle to create a complete heart block model. The heart rates of pigs in both groups were observed after isoproterenol （ISO） being injected through femoral vein. Results： The isolated cells had good viability and all the pigs showed complete heart block ECG waveforms after ablation. Ventricular autonomic rhythms of pigs in transplantation group were faster than those of pigs in control group （P〈0.05）. We proved that the rhythms originated from the injected site by endocardial pace mapping. The ventricular rhythms of transplantation group changed remarkably after ISO was administrated （P〈 0.05）. Conclusion： Autogeneic transplantation of SAN cells isolated from young pigs into right ventricular wall adjacent to heart apex can increase the ventricular autonomic rhythm of complete heart block, and this increased rhythm is sensitive to isoproterenol.[

Abstract:Objective..To prepare a novel surgery mesh made of polydioxanone（PDO） and to study its in vivo degradation properties and the related tissue reaction, so as to provide reference for future clinical application. Methods： PDO mesh was plain-woven and was coated with collagen and chitosan, then they were implanted into the dorsal muscle of rabbits for 2, 4, 8, 12, 16, and 24 weeks. Rabbits implanted with pure PDO mesh served as control. The degradation process of implanted materials, tissue-material interface, and the biological influence of the coating on the mesh were observed; tissue reaction was observed and evaluated by histological means and TEM. Results.. Implantation of pure PDO mesh resulted in mild inflammation and fibrous tissues infiltrated into the mesh; degradation occurred during the 8^th to the 12^th week and degraded completely within 24 weeks; there was no denaturalization, necrosis, and abnormal hyperplasia in the surrounding tissues. Collagen coating stimulated a fibroblastic proliferation and was reabsorbed within 4 weeks ; and the naked mesh was infiltrated by fibrous tissues. The chitosan coating showed a strong inhibitory effect on the growth of the surrounding connective tissues. Conclusion： The biodegradable polydioxanone mesh has an excellent biocompatibility, a friendly tissue-material interface and a proper degradation rate matching to the tissue regeneration rate. The biological characteristics of the mesh can be modified by different coatings, making it a very promising degradable surgical mesh.[

Abstract:Objective： To prepare amphotericin B-polybutylcyanoacrylate-loaded nanoparticle （AmB-PBCA-NP） and to assess their ability to pass the blood-brain barrier. Methods.- AmB-PBCA-NP was prepared by incubation, with the surface modified by polysorbate 80. High-performance liquid chromatography （HPLC） method was established using acetonitrile and water （40 ： 60, 4% of acetic acid） as mobile phase; detection was done at 405 nm. Mice were divided into 3 groups： AmB- injected, AmB-lipid-injected, and AmB-PBCA-NP-injected group. Brain-targeting ability of AmB-PBCA-NP was then evaluated by determination of AmB concentrations in mice brain and other specimens. Results： The mean diameter of the prepared AmB- PBCA-NP was 94.38 nm; the mean entrapment efficiency of the particles was 82% ; and the drug loading rate was 56.10%. It was found that AmB alone failed to pass the blood-brain barrier; only very low level of AmB was detected in AmB-lipid group 3 h after injection; moderate concentration of Amb was detected 30 min after injection of AmB-PBCA-NP and the concentration peaked at 133 ng/g 3 h after injection. Conclusion： AmB-PBCA-NP modified by polysorbate 80 can pass the blood-brain barrier

Abstract:Objective：To evaluate the in vitro release behavior of doxorubicin（Dox）-loaded microspheres and the stability of Dox during encapsulation process and in vitro release. Methods.. Dox-loaded microspheres were prepared by double emulsion （W/O/W） method with poly（lactic-co-glycolic acid） （PLGA） as the carrier material. The physical and chemical characteristics of mierospheres, including the mean diameter, morphology, drug entrapment efficiency and loading rate, were evaluated. The in vitro release behavior and its influencing factors were determined by ultraviolet spectrophotometry. Dox stability was evaluated by HPLC method during the encapsulation process and in vitro release. Results： The prepared microspheres had a complete spheric shape and dispersive quality. The mean diameter of the microspheres was 85 μm; the drug entrapment efficiency was 95.1%; and the loading rate was 14.8%. Releasing rate of the microspheres slowed down with the increase of PLGA concentration and the decrease of W/O value. The encapsulation process had no obvious effect on the stability of Dox, while Dox degraded during in vitro release as the prolongation of time. On day 10, the peak area of degraded material accounted for 2.46%. Conclusion： Dox can be encapsulated in the microspheres by double emulsion method and different release rates of Dox can be achieved by adjusting PLGA concentration and W/O volume ratio.[

Abstract:Objective：To study the influence of anti-tumor drug incorporation on the physicochemical properties of calcium phosphate cement（CPC）. Methods： Methotrexate （MTX）, epirubicin （EPI）, hydroxy camptothecin （OH-CPT）, and arsenic trioxide （As2O3） were incorporated, each in a proportion of 2%, 5 %, and 8%, into the powder-phase CPC. Untreated CPC was taken as control. The setting time, compression strength, and the microstructure of the resultant products were evaluated and tested. Results： Compared with control group, the setting time was significantly prolonged when 2 % EPI was incorporated into CPC （P〈0.05）, and the maximum compression strength was significantly decreased when 5% OH-CPT and 8% MTX, EPI were incorporated into CPC （P〈0.05）. Electron microscope showed no significant change in the crystal structure after incorporation of the drugs. Conclusion： CPC is an ideal anti-tumor drug delivery material. The changes of physicochemical properties of CPC must be considered when anti-tumor drugs are incorporated into CPC for clinical application.[

Abstract:Objective：To explore the role of reactive oxygen species （ROS,i. e, H2O2 and O2^- ） in regulation of respiratory rhythm in the medial area of nucleus retrofacialis （mNRF）. Methods： Medullary slices of neonatal SD rats, including hypoglossal nerve （Ⅻ n） and mNRF, were made according to Suzue＇s method. Simultaneous recording of the Ⅻ n respiratory rhythmic activity （RRA） with suction electrode and the respiratory neuronal discharge were performed with whole cell patch in the mNRF on the brainstem slice in vitro. The effect of t-butyl hydroperoxide （tBHP） and α-lipoic acid （α-LA） on the respiratory pacemaker neurons and respiratory rhythm in the mNRF were observed. Results： tBHP significantly decreased respiratory cycle （RC） and increased respiratory amplitude; α-LA significantly increased RC and decreased its amplitude. Meanwhile, α-LA significantly prolonged the action potential of the respiratory cadmium-insensitive pacemaker neurons and reduced its amplitude, but it had no significant effect on the cadmium-sensitive respiratory pacemaker neurons. Voltage steps and ramps showed that α-LA inhibited both the transient and persistent sodium current of cadmium-lnsensitive pacemaker neurons. Conclusion： ROS has an excitatory effect on the respiratory rhythm and the cadmium-insensitlve respiratory pacemaker neurons through modulating sodium current.[

Abstract:Objective：To construct Mycobacterium tuberculosis ESAT6 （pE） antigen DNA vaccine and ubiquitin-ESAT6 fusion gene （pUE） DNA vaccine. Methyls： The constructed DNA vaccines were intramuscularly inoculated into female BALB/c mice separately. The serum antibodies （including IgG, IgG1, and IgG2a ）, Cytokines （IFN-γ and IL-4）, and cytotoxic T lymphocytes response were detected in the immunized mice. Results： Mice in pE group had a higher serum level of IgG （P〈 0.01） and a lower value of IgG2a/IgG1 （[2.28±0.40] vs [3.87±0.60] ,P〈0.05） than mice in pUE group. Besides, mice in pUE group secreted more IFN-γ than those in pE group （P〈0.01）, but secreted less IL-4 （P〈0.01）. Furthermore, pUE enhanced the activity of CTL. The results showed that pUE DNA vaccine induced weaker humoral immune response, but stronger cellular immune responses compared to pE DNA vaccine. Conclusion.. The pUE DNA vaccine constructed in this study sheds new lights on the prophylactic and therapy of tuberculosis.[

Abstract:Objective：To study the isolation, identification, and biological characteristics of the secondary metabolites of myxobacteria. Methods： A growth inhibition model of Pyricularia oryzae was used to screen the active microbes. The compounds extracted with methanol from the fermentation broth of Myxococcus xanthus 095B06 were separated by silica chromatography, gel filtration chromatography, and high-performance liquid chromatography. The chemical structures of the compounds were identified by ^1HNMR, ^13CNMR, ESI-MS, and EI-MS techniques. The hioactive activities of the separated compounds were evaluated by Kirby-Bauer Disc Diffusion method and MTT method. Results： Six compounds, namely, Avermectin Ala, Avermectin A2a, Avermectin Bla, Avermectin B2a, ergosta-7,22-dien-3,5,6-triol, and 4-quinolinecarboxylic acid, were obtained. Conclusion： Compound 1, 2, 3, 4, and 5 have been isolated from myxobacteria for the first time and compound 2 and 4 can strongly inhibit the growth of SMMC-7721 cell line, with both IC50 values being 5 g/ml.[

Abstract:Objective： To investigate the influence of trichostatin A （TSA）, a histone deacetylase （HDAC） inhibitor, on the growth of human bladder cancer cells and on the expression of related genes, and to explore the mechanism involved. Methods： MTT assay was employed to evaluate the inhibitory effect of TSA（0. 05, 0. 1, 0.2, 0. 4, 0. 8 μmol/L）on growth of human T24 bladder cancer cells. The morphological changes of T24 cells were observed by transmission electron microscope after treated with 0. 4 μmol/L TSA ; the cell cycle distribution and apoptotic ratio were determined by flow cytometry. The acetyl level of histone after TSA treatment was detected by Western blot; the rnRNA expression of p21^CIP1/WAF1 , cyclin A, and cyclin E was measured by FQ-PCR. Results： MTT assay revealed TSA inhibited the growth of T24 cells in a concentration- and time-dependent manner. Typical morphological changes of apoptotic cells were observed by electron microscope after treatment with 0.4 μmol/L TSA. Flow cytometry showed that the cells were blocked at Go/G1 phase and typical Sub-G1 peak appeared. TSA obviously promoted the acetyl level of histone, induced expression of p21^CIP1/WAF1 mRNA, and inhibited expression of cyclin A, but had no obvious influence on expression of cyclin E. Conclusion： TSA can inhibit bladder cancer cells through inducing cell apoptosis and cell cycle arrest in vitro, which might be related to the acetyl level of histone and the expression of p21^CIP1/WAF1 and cyclin A.[

Abstract:Objective：To investigate the protective effects of Er-xian decoction on ovariectomy-induced osteoporosis in rats. Methods.. The osteoporosis model was induced by ovariectomy in rats. Twelve weeks later, the undecalcified longitudinal proximal tibial metaphysical sections were made and stained for the bone histomorphometric analysis. Bone density of femur metaphysis was determined by dual-energy X-ray absorptiometry. Results： Er-xian decoction significantly increased trabecular area and trabecular thickness （P〈0.01） and decreased trabecular separation（P〈0.05）. The parameters of bone formation of rats, such as MAR, BFR/BS, BFR/BV, and BFR/TV, were increased significantly after Er-xian decoction treatment（P〈 0. 05, P〈0.01）. Moreover, Er-xian decoction obviously reduced osteoclast number（P〈0.01）and enhanced bone density of femur metaphysis in ovariectomized rats （P 〈 0. 05 ）. Conclusion： Er-xian decoction has antiosteoporotic effects on ovariectomized rats by promoting bone formation, increasing bone density, and inhibiting bone resorption.[

Abstract:Objective.. To study the protective effect of Tongluo recipe on renal function of rats with diabetic nephropathy and its influenee on the oxidative stress and antioxidative system in renal cortex. Methods.. Diabetes mellitus was induced in male SD rats by intraperitoneal injection of streptozotocin（55 mg/kg）. Rats with a blood glucose level ≥16. 7 mmol/L after 3 days were taken as diabetic rats. Twenty-one diabetic rats were evenly randomized into 3 groups, namely, untreated diabetic control （DM group）, Tongluo recipe garaged （1 g · kg^-1· d^-1 for 12 weeks, TLR group）,and melatonin garaged （5 mg· kg^-1· d^-1 for 12 weeks, MEL group） groupsl normal rats were taken as normal control （NC group）. The treatments were started 3 days after streptozotocin injection and continued for 12 weeks. Kidney cortex was collected under anesthesia for electron microscopic observation. the kidney homogenates were prepared to determine the renal anti-oxidant enzyme（SOD, GSH-Px） activities and lipid proxide （MDA） levels by chromatometry. Results.. Compared with NC group, rats in DM group had a higher 24 hour-urinary protein excretion （P〈0.01）, while those in TLR group and MEL group were significantly lower than that of DM group （P〈 0. 01 ）. Electron microscopic results showed that the rnesangial ceils and endothelial ceils in DM group were swollen and had fusion/disappearance of podocytic process, increased mesenteric matrix, uneven basal membrane, and increased mitochondria with morphological disorders. while the above changes were improved in TLR and MEL group. GSH-Px and SOD activities were lower and MDA levels were higher in renal cortex of DM group than those of NC（P〈0.05） . treatment with Tongluo recipe and melatonin slightly increased SOD and GSH-Px activities and decreased MDA levels, with the level in MEL group significantly lower than that of DM group （P〈0.05）. Conclusion： Tongluo recipe can protect the renal function of diabetic rats by lowering urinary protein excretion and improving the structure of glomerular basement membrane, but it can only slightly decrease MDA level in the renal cortex, showing an unclear anti-oxidative effect.[

Abstract:Objective：To investigate the anti-fungal activities of 20 tetralin compounds in vitro. Methods： We adopted the M27-A project recommended by the National Committee for Clinical Laboratory Standard （NCCLS）. Tetralin compounds were tested and selected with several candidal strains and non-candidal strains with different susceptibilities to fluconazole. After obtaining the susceptibility results, we plotted the time-growth curves of several typical tetralin compounds, including compound 22-1, 31-1 and their muriate 22,31, as well as compound 34-1,26-1 and their bromate 34,26. The combination effects of compound 22, 26, 34, 31-1 with other anti-fungal agents （with different structures） were also determined. Results： The 20 tetralin compounds were proven to have different degrees of anti-fungal activities. Compound 31-1 had a stronger anti-fungal activity to FLC-susceptible strains than fluconazole did, and its effect lasted for over 54 h at the concentration of 6 μg/ml. Compounds 22-1 and 22 markedly suppressed the growth of Candida albicans, Candida parapsilosis, and Trichophyton rubrum, with all the MIC50 values less than 0. 125μg/ml. Time-growth curves indicated that the anti-fungal activity of 22-1 on fluconazole-resistant strain was more powerful than that of ketoconazole. Furthermore, tetralin compounds had a synergistic effect with terbinafine and berberine on fluconazole-resistant strains. Conclusion： Tetralin compounds have powerful anti-fungal activities and their structures are different from those of other anti-fungal agents currently used, which provide a basis for developing new anti-fungal agents.[

Abstract:Objective：To investigate the chemical constituents in the roots of Angelica dahurica var. formosana. Methods： The dried rhizomes of Angelica dahurica var. formosana were chopped and extracted with 80% EtOH for 3 times under reflux. The EtOH extract was isolated by chromatography on silica gel using a gradient solvent system （PE： EtOAc ＆ CHCl3 ： MeOH） and Sephadex LH-20. The structures of the contents were elucidated by nuclear magnetic resonance and MS spectral analysis. Results： Seventeen compounds were identified, namely, isoimperatorin （ 1 ）, imperatorin （ 2 ）, bergapten （ 3 ）, alloimperatorin（ 4 ）, xanthotoxol （ 5 ）, cnidilin （ 6 ）, byakangelicin （ 7 ）, bergaptol （ 8 ）, byakangelicol （ 9 ）, pabulenol （ 10 ）, oxypeucedanin hydrate（ 11 ）, desmodimine （ 12 ）, palmitic acid （ 13 ）, stiamasterol （ 14 ）, β-sitosterol （ 15 ）, β-daucosterin （ 16 ）, and sucrose（17）. Conclusion： Compounds 8 and 12 have been isolated from Angelica dahurica var. formosana for the first time. Compound 12 has been firstly isolated from Umbelifera plants. All the courmarin compounds belong to linear 6, 7- furanocourmarins.[

Abstract:Objective： To establish a liquid chromatography-mass spectrometry （LC-MS） method for determining the concentration of amlodipine besylate in human plasma and to evaluate the bioequivalence of 2 kinds of amlodipine besylate tablets. Methods： Twenty healthy male volunteers were enrolled into a single crossover study. A single dose of the suspension equivalent to 10 mg amlodipine besylate or a reference preparation was given in a crossover way. The plasma concentrations of amlodipine besylate were determined by LC-MS method in the volunteers at different time points; the pharmacokinetic parameters and relative bioavailability were calculated and the bioequivalence of the 2 preparations were evaluated. Results： The pharrnacokinetic parameters for experimental and the reference preparations were： Cmax （6. 21 ± 1. 88） vs（ 6. 03 ± 1. 08） ng/ml; AUC0-120（250. 68±52. 61） vs （246.14±52.11） ng h/ml; Tmax（6.0±2.3） vs （6. 1± 2.5） h; t1/2（40.45±6.68） vs （43.74±9.05） h, respectively. The linear range of the present method was 0. 1-20. 0 ng/ml; the lowest detectable concentration of amlodipine besylate was 0.1 ng/ml. There was no significant difference in pharmacokinetic parameters between the 2 tablets. Conclusion： The present method is simple to use, fast, and accurate. The 2 preparations of amlodipine besylate are bioequivalent.[

Abstract:Objective：To examine the expression of α-melanocyte-stimulating hormone（α-MSH） and the melanin in different kinds of human skin autografts and in the normal skins, so as to elucidate the role of α-MSH in hyperpigmentation in the skin autografts. Methods： Immunohistochemical technique was employed to detect the expression and distribution of α-MSH in skin autografts （including the full thickness skin autografts, medium thickness skin autografts, razor-thin skin autografts, and normal skins adjacent to the donor site and the recipient site）. Masson-Fontana staining technique was used to detect the melanin contents in all the above skin specimens. Results： The location of α-MSH expression was at the cytoplasm of melanocytes and keratinocytes in epiderm; α-MSH was positive in most skin autografts and its expression was higher in the thinner skin autografts. The expression of α-MSH in all types of skin autografts was significantly different from that in normal skin （P〈 0.01） ; α-MS expression was also significantly different between all the skin autografts （P 〈 0. 01） ; α-MSH expression in normal skin around donor site and recipient site autografts was obvious increased compared with that in normal had no statistical difference. The contents of melanin in skin skin（P〈0.01） ; the contents of melanin between all the skin autografts were also significantly different （P〈0.01）. The melanin contents increased with the decrease of skin autografts thickness. The expression of α-MSH was positively correlated with the contents of melanin in epidermis. Conclusion： The expression of α-MSH in skin autografts is positively correlated with the contents of melanin in skin autografts. Overexpression of α-MSH may play an important role in hyperpigmentation process of skin autografts.[

Abstract:Objectlve：To assess the clinical outcome of autologous fat injection in treatment of hemifacial atrophy, so as to search for an ideal treatment of hemifacial atrophy. Methods： Autologous fat granules were obtained by tumescent liposuction. After purification, the fat granules were injected in a multi-strata, multi-tunnel, and multi-spot manner to repair facial introcession abnormality. Patients whose introcession was not improved after the stabilization of the fillings were re-injected with autologous fat granules twice or more until satisfactory outcomes were achieved. Results： All the 7 patients in this study obtained satisfactory outcomes after fat injection and basically reached symmetry. The follow-up result was good after 2 years. Conclusion： Local injection transplantation of autologous fat granules is effective for the treatment of facial depression and hemifacial atrophy, with slight trauma, little complication, and satisfactory outcome.[

Abstract:Objective..To investigate the morphological changes of cerebral blood vessels by digital subtraction angiography （DSA） in patients with central vertigo, so as to provide references for diagnosis and treatment of patients with central vertigo. Methods： Ninety-two patients suffering from central vertigo were subjected to cerebral angiography through femoral artery. The types, proportion, and distribution （site predilection and number of involved vessels） of the cerebrovascular abnormalities were observed and analyzed. Results： Of the 92 patients with central vertigo, cerebrovascular lesions were found in 137 sides, including 15 （10.95 % ） anterior circulation lesions and 122 （89.05% ） posterior circulation lesions. Thirty-one （ 33.7 % ） patients had 2 or more lesions. The most common lesion was vertibroarterial lesion accompanied with other cerebrovascular stenosis. The lesions included 39 （28.47 %） sides with vertibro-basilar stenosis or occlusion, 13 （9.49 %） sides with posterior cerebral artery stenosis or occlusion, and 59 （43. 06%） sides with posterior circulation abnormalities. Conclusion： The posterior eireulatlon stenoses or abnormalities might be the major reasons for central vertigo in middle- and old-age patients.[

Abstract:Objective： To analyze the influencing factors of investment management of hospital intangible assets, so as to provide countermeasures for establishing and optimizing the investment management system of hospital intangible assets. Methods： Taking hospital reputation as an example, we analyzed its role in the investment management of hospital intangible assets and the maintenance of hospital reputation in an economic perspective using Shapiro model and game theory. Results： The factors, such as the blueprint of hospital, efficiency of medical quality information transmission, and cost variance, all influenced the reputation of hospital. Conclusion： In the health care field, stable property right system should be established and market information agents should be cultivated. The administrative actions of involved departments should abide by the related regulation and the running system of hospitals should be optimized.[

Abstract:Objective： To investigate the in vivo absorption kinetics of caffeine at different intestine segments in rats. Methods： The rat intestine was cannulated for in situ recirculation. The absorption kinetics of caffeine,including the absorption segments of intestine,drug concentration and p H value,were investigated in all the rats under light anesthesia. Ultraviolet light spectrometry and high-performance liquid chromatography were used to determine the concentrations of phenol red and caffeine, respectively. Results： The absorption rate constants （Ka） of caffeine in the small intestine at the concentrations of 2.5,5,10 and 25 mg· L^-1 were0.252,0.247,0.304 and 0.232 h^-1,respectively. The Ka values at pH of 7.8,6.8,and 5.4 were 0.267, 0. 274 and 0. 247 h^-1 ,respectively. The Ka values in the duodenum,jejunum,ileum,and colon were 0. 112,0. 099,0. 095 and 0. 069 5 h^-1 ,respectively. Conclusion, Concentration and pH value have no obvious effect on the absorption kinetics of caffeine. Caffeine can be better absorbed in the duodenum,jejunum and ileum than in the colon. The absorption of caffeine in the intestine is a first-order process through passive diffusion mechanism.[

Abstract:Adult stem cells have great advantage in tissue reconstruction and regeneration. Transplantation of autologous stem cells into ischemic tissue is a novel therapeutic option for ischemic disorders. This review summarizes the potential role of adipose tissue-derived stromal cells （ADSC） on regenerative cell therapy for ischemic diseases. ADSC can be readily harvested and cultured, under specific condition,they can be induced to differentiate into adipocytes, bone, neurons,and endothelial cells. Moreover,ADSC can secrete a number of angiogenesis-related cytokines which might be suitable for regenerative cell therapy. It has also been reported that ADSC could differentiate into myocardlocytes. ADSC might be an important material for regenerative cell therapy in the near future,replaclng bone marrow cells.[

Abstract:Unfolded protein response（UPR） is a protective response in cell endoplasmic reticulum （ER） under stress condition. Three ER transmembrane proteins, IRE1, PERK, and ATF6, coordinately regulate the UPR function in mammalian cells through their signaling pathways. In addition, some proteins and transcription factors during the UPR can provide negative and positive feedback loops to maintain the normal function of ER. UPR can trigger cell death or apoptosis and eventually cause related diseases if the ER stress persists. Several key mediators of UPR are candidates for therapeutic targets in many studies. Up to now progress has been made in the area, which provides new ideas for clinical practice and holds a great potential for future application.[

Abstract:Basic fibroblast growth factor（bFGF） is one of the most important factors for wounds healing. BFGF promotes healing of bone fracture by regulating cell proliferation and differentiation of bone tissues, increasing local bone density, and accelerating local angiogenesis. With the progression of bFGF research, more and more attention will be paid to bone repair function of bFGF in bone tissue engineering.[

Abstract:高等动物的基因表达分析往往受制于大量纯化同质细胞的取材困难,由于细胞异质性的影响,常规研究方法也无法识别细胞间表达特性的差异。单细胞水平是切实表明综合性基因表达最行之有效的研究途径,而单细胞逆转录PCR又是该水平上基因表达研究最有力的工具。与常规RT-PCR相比,该技术在各个环节上都有着显著差异。为此,详尽论述了单细胞逆转录PCR技术在各个环节上的要点包括显微操作、扩增和分析方法、基本注意事项等内容以及该技术在实际科研中的应用情况。[

Abstract:目的：运用经胸超声心动图（TTE）观察冠心病患者冠脉介入治疗前、后冠状静脉窦（CS）的血流变化,以探讨超声测定冠状静脉窦血流用于评价冠脉介入治疗后冠脉灌注改善情况的可能性。方法：22例拟行经皮冠状动脉内成形术（PTCA）和内支架置入术（ICS）的冠心病患者（皆为左冠状动脉及其属支病变）,包括单部位介入（11例）、双部位介入（9例）和三部位介入（2例）。治疗前后分别行经胸超声心动图检查,测量及计算冠状静脉窦的指标为：平均内径（Dmean）、平均流速（Vmean）、速度时间积分（VTI）、血流量（Q）;同时测定左室射血分数（LVEF）、二尖瓣前向血流指标（E、A及E/A）等心功能指标。结果：在冠脉介入术后,左室心功能指标均较术前明显改善（P〈0.05）;冠状静脉窦的Vmean、VTI和Q均有显著增加（P〈0.05）。结论：TTE测定冠状静脉窦血流可以反映PTCA及ICS术后冠脉灌注的改善情况。[

Abstract:目的：评价科玛嘉念珠菌显色培养基在酵母菌临床鉴定中的应用价值。方法：临床标本先用沙堡培养基培养,分离出的酵母菌再用科玛嘉念珠菌显色培养基鉴定。结果：8525份临床标本培养阳性2687例,阳性率为31.52%,其中酵母菌2485株,通过科玛嘉显色培养基鉴定到种的有2188株,占88.11%。结论：科玛嘉念珠菌显色培养基能快速、有效地鉴定临床常见的4种酵母菌。[

Abstract:目的：探讨长春新碱载体红细胞对昆明小鼠S180肿瘤的抑瘤作用。方法：用昆明种小鼠右侧腋窝皮下接种S180肉瘤细胞建立荷瘤小鼠模型,将48只荷瘤小鼠随机分为长春新碱组、长春新碱载药红细胞组、未载药红细胞组和阴性对照组4组,每组各12只。分别经尾静脉注射长春新碱、长春新碱载药红细胞、洗涤红细胞及生理盐水,1次/3 d,共5次。所有治疗结束次日处死小鼠并称重,称瘤质量,计算各组瘤体平均质量及肿瘤抑瘤率。结果：长春新碱载药红细胞组的体质量增长幅度为（36.31±1.51）g,明显大于长春新碱组的（31.32±3.55）g（P〈0.01）。长春新碱载药红细胞组平均瘤质量最轻,为（0.33±0.05）g,明显低于阴性对照组的（0.57±0.05）g（P〈0.05）,该组抑瘤率达42.42%,是长春新碱组17.49%的2.43倍;未载药红细胞组瘤质量较阴性对照组大,抑瘤率呈负值,为-13.76%。结论：长春新碱载体红细胞体内抗肿瘤活性显著,有较强抑瘤作用,为临床肿瘤治疗提供新思路和可行方法。[

Abstract:目的：制备盐酸哌甲酯缓释微丸,并对其体外释药情况进行研究。方法：采用流化床包衣技术制备盐酸哌甲酯缓释微丸。对包衣材料种类、配比及用量进行选择,建立HPLC法测定包衣微丸体外释放度。HPLC色谱条件：色谱柱：Diamonsil C18（5μm,200 mm×4.6 mm）;流动相：甲醇-0.02 mol/L磷酸二氢钾-冰乙酸（40：55：5）;检测波长：210 nm;流速：0.8 ml/min;进样量：20μl。结果：乙基纤维素水分散体（Surelease）比丙烯酸树脂水分散体（Eudragit NE30D）更适合作为盐酸哌甲酯缓释微丸的包衣材料。包衣增重15%,HPMC加入量为6%时包衣微丸在不同释放介质（水、pH 1.2盐酸溶液及pH 4.0、pH 6.5p、H 6.8、pH 7.5磷酸盐缓冲溶液）中均呈现良好的缓释效果。结论：成功地制备了盐酸哌甲酯缓释微丸,其体外释药缓慢、持续、平稳。[

Abstract:近年来介入封堵先天性心脏病室间隔缺损（VSD）特别是膜部缺损，具有疗效确切、创伤小、并发症少、不需用体外循环等优点，已成为治疗的首选，但对于嵴内型VSD，因其解部位置特殊，邻近瓣膜组织，部分病例可因缺乏支撑而致主动脉瓣脱垂于缺损内并伴有主动脉瓣关闭不全，而封堵后也有可能引起瓣膜功能不全，是否行介入封堵尚有争议。我们应用国产零偏心封堵器成功封堵7例嵴内型VSD，近期效果良好，现报告如下。[

Abstract:自1976年Trager和Jensen成功建立了恶性疟原虫体外连续培养方法以来，该方法已广泛应用于恶性疟原虫分子生物学及免疫学的研究。恶性疟原虫的冻存与复苏，是能否成功进行体外培养的关键。国内外研究者对寄生虫冻存方法的探索已持续了半个多世纪，但就如何最大限度地减少低温对寄生虫的损伤，仍未得到规律性的结论。本实验对几种恶性疟原虫冻存复苏的方法进行了探讨，以期得到提高恶性疟原虫冻存后复苏成活率的更为有效的方法，为恶性疟原虫的体外培养提供有力的支持。[

Abstract:心律失常的射频消融治疗目前已得到广泛的开展。房性心动过速（房速）作为一种常见的心律失常，也是射频消融的良好适应证。其发生与心房的特殊解剖结构密切相关，并且不同部位起源的房速，在发生机制、诱发、标测及消融方面均有各自的特殊性。右房界嵴部位解剖结构特殊，是房速的好发部位。我们成功消融了15例界嵴部位的房速，现对房速的电生理特点及其射频消融治疗作一总结。[