Cytoplast and microprotoplast are main subprotoplasts that can play an important role in plant genetic improvement.The present review highlights the advancements in isolation and fusion of plant subprotoplast,and some suggestions and prospects are proposed for the future studies.

Objective To explore the abilities and personal qualities that medical undergraduates need for their future clinical work, and develop their competency model. Methods The self-made ques-tionnaire survey on medical undergraduates' competencies was conducted among the 1326 medical under-graduates in a medical university by using the cluster sampling method, and the 1099 effective question-naires were collected with effective rates of 82.88%. The exploratory factor analysis was used to analyze the internal structure of competency item and to build the competency model. The rating method was used to calculate the weight of each competency item. Results The overall Cronbach's alpha coefficient of the competency questionnaire was 0.965, and the coefficient of each dimension was above 0.832. The KMO statistic value was 0.970, and the probability associated with Bartlett test statistic was P=0.000, showing that the questionnaire has high reliability and validity. The competency model of medical undergraduates covers six aspects, namely, craftsmanship, personal qualities, professional learning, interpersonal communication, psychological adaptation and the pursuit of excellence, which contain 49 factors. The cumulative variance contribution rate was 54.729%. The weights of these six aspects in the model were 0.204 4, 0.202 1, 0.175 3, 0.158 8, 0.137 6 and 0.121 8 respectively. Conclusion The competency model of undergraduate medi-cal students has a certain scientific and practical value, which can provide new evaluation methods and ideas for medical education objectives, quality assurance, teaching evaluation, medical personnel selection and training.

Background Previous experimental and clinical studies have proved that elevated serum uric acid increased risk for developing hypertension.Whereas,there are a paucity of information on the relationship between serum uric acid and prehypertension.Objective The purpose of this research is to evaluate the association between the serum uric acid and prehypertension.Method A cohort of seven thousand eight hundred thirty-nine subjects without hypertension and other cardiovascular diseases were recruited from a cross sectional study in urban and rural place in 9 provinces during 2005-2006.Based on serum uric acid(324 ?mol/L for overall population,366 ?mol/L for male,285 ?mol/L for female),people were categorized into quartiles.The odds ratio for prehypertension was calculat ed with the lowest quartile as the reference.Results The prevalence of prehypertension increased with increasing uric acid in total population(P324 ?mol/L)to lowest quartile 1(

Objective To establish high-performance liquid chromatography with electrochemical detector(HPLC-ECD) method for the determination for metanephrine and normetanephrine in 24 h urine, and provide a superior test for the diagnostic of pheochromocytomas over plasma/urine catecholamine.Methods MCX solid-phase cartridge was used for extraction of metanephrine and normetanephrine,HPLC-ECD was used for their measurements.The intra-assay CVs,interassay CVs and recoveries of metanephrine and normetanephrine were also calculated.104 hypertensive patients without pheochromocytomas and 5 pheochromocytomas patients were selected in this study.The concentrations of metanephrine and normetanephrine were compared with the plasma and 24h urinary catecholamines concentrations.Results The intra-assay CV,inter-assay CV and recovery of metanephrine were 5.9%, 7.5%,91.1% respectively;the intra-assay CV,inter-assay CV and recovery of normetanephrine were 6.3%,6.6%,88.5%,respectively.The MN,NMN,plasma CA and urine CA of all pheochromocytomas patients were positive.MN and NMN were negative in controls,while plasma CA and urine CA are false positive in 15 patients and 14 patients in controls,respectively.Conclusions The study establish a fast and accurate method for quantification of metanephrine and normetanephrine in 24 h urine by HPLC-ECD.These findings also prove that it is the best biochemical assays for pheochromocytomas at present.

Objective To study the value of fast dynamic contrast enhanced MRI in the diagnosis of orbital occupying lesions. Design Retrospective ease series.Participants 21 patients with orbital occupying lesions.Methods All the 21 patients were performed on fast dynamic contrast enhanced MRI and were verified by pathology.The raw datum were processed by the software of GE Functool. Parameters such as time-intensity curve(TIC),time to peak(Tpeak),1 minute enhancement ratio(ERlmin)and maximum enhancement ratio(ERmax)were analyzed to study the characteristics of orbital diseases on fast dynamic contrast-enhanced MRI.Main Outcome Measures TIC,ERlmin,and ERmax.Results The characteristics of TIC between benign diseases and malignant tumors were different. Of the 16 cases of benign lesions,12 demonstrated as continuous increasing type,and of 5 cases of malignant lesions,3 cases of lym- phoma were all platform type.The ER1min of the malignant tumors(150.47?42.18)was higher than that of the benign lesions (101.37?43.02)(P=0.021).Cavernous hemangiomas had special progressing enhancing model.Conclusions Fast dynamic contrast enhanced MRI is valuable to distinguish malignant tumors from benign occupying lesions.(Ophthalmol CHN,2007,16:305-308)

OBJECTIVETo compare the differences in the efficacy on distant version of naked eye in the patients of juvenile myopia between rotating manipulation and lifting-thrusting manipulation of acupuncture.

METHODSOne hundred and twenty cases (240 eyes) were randomized into a rotating manipulation group and a lifting-thrusting manipulation group, 60 cases (120 eyes) in each group. Additionally, a corrective lenses group, 60 cases (120 eyes), was set up as the control. In both manipulation groups, Cuanzhu (BL 2),Yuyao (EX-HN 4), Sizhukong (TE 23), Taiyang (EX-HN 5), Fengchi (GB 20), Zusanli (ST 36), Guangming (GB 37) and Sanyinjiao (SP 6) were punctured, but stimulated with rotating manipulation and lifting-thrusting manipulation respectively three times per week, 10 times as a treatment session and totally one session was required. In the corrective lenses group, the glasses were applied at daytime. The clinical efficacy and the changes in distant vision of naked eye before and after treatment were compared among the three groups.

RESULTSThe total effective rate was 87.5% (105/120) in the rotating manipulation group, which was better than 69.2% (83/120) in the lifting-thrusting manipulation group (P < 0.05). The distant vision of naked eye was improved apparently in the rotating manipulation group and the lifting-thrusting manipulation group after treatment (both P < 0.05). But it was not improved in the corrective lenses group (P > 0.05). The distant vision of naked eye was improved more apparently after treatment in the rotating manipulation group as compared with that in the lifting-thrusting manipulation group (0.75 +/- 0.23 vs 0.68 +/- 0.24, P < 0.05). For 96 cases (192 eyes) with acupuncture treatment, in 3-month follow-up, 87.0% (167/192) of the cases maintained the stable vision as the original level and 13.0% (25/192) of them were reduced in the vision In the acupuncture groups, it was found that the improvement of distant vision of naked eye was more obvious after treatment with younger age, better basic vision and shorter duration of sickness (all P < 0.05).

CONCLUSIONAcupuncture achieves the positive and sustainable clinical effect on juvenile myopia, and the results of rotating manipulation are superior to that of lifting-thrusting manipulation. Age, basic vision and duration of sickness impact the clinical efficacy.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Adolescent ; Child ; Female ; Humans ; Myopia ; therapy ; Young Adult

Analysis of common volatile constituents in herbal pair (HP) Chuanxiong Rhizome (CXR)-Notoperygium root (NR) and its single herb was performed by the method of alternative moving window factor analysis (AMWFA). In total, 65, 71, and 79 volatile chemical components in volatile oil of CXR, NR, and HP CXR-NR were separately determined qualitatively and quantitatively, accounting for 83.69%, 96.04% and 95.39% total contents of volatile oil of CXR, NR, and HP CXR-NR respectively. Analysis by the method of AMWFA indicates that there are 45 common volatile constituents in HP CXR-NR and single herb CXR, and 63 common volatile constituents in HP CXR-NR and single herb NR and 31 common volatile constituents among these three systems. The experimental results also show that the number of volatile chemical components in HP CXR-NR is almost equal to the sum of the number in the two single herbs, and volatile chemical components in HP CXR-NR are mainly from single herb NR.
Asteraceae ; chemistry ; Gas Chromatography-Mass Spectrometry ; Ligusticum ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry

OBJECTIVETo investigate telomerase activity in rabbit bone marrow stromal cells (BMSCs) during their committed differentiation in vitro along neural pathway and the effect of glial cell line-derived neurotrophic factor (GDNF) on the expression of telomerase.

METHODSBMSCs were acquired from rabbit marrow and divided into control group, GDNF (10 ng/ml) group. Cytokine.NSCs medium (prepared by our lab, Patent No. ZL02134314. 4) supplemented with 10 percent fetal bovine serum (FBS) was used to induce BMSCs differentiation along neural pathway. Fluorescent immunocytochemistry was employed to identify the expressions of Nestin, neuron-specific endase (NSE), and gial fibrillary acidic protein (GFAP). The growth curves of the cells and the status of cell cycles were analyzed, respectively. During the differentiation, telomerase activities were detected using the telomeric repeat amplification protocol-enzyme-linked immunosorbent assay (TRAP-ELISA).

RESULTSBMSCs were successfully induced to differentiate along neural pathway and expressed specific markers of fetal neural epithelium, mature neuron and glial cells. Telomerase activities were undetectable in BMSCs during differentiation along neural pathway. Similar changes of cell growth curves, cell cycle status and telomerase expression were observed in the two groups.

CONCLUSIONSRabbit BMSCs do not display telomerase activity during differentiation along neural pathway. GDNF shows little impact on proliferation and telomerase activity of BMSCs.

Animals ; Bone Marrow Cells ; enzymology ; Cell Differentiation ; Glial Cell Line-Derived Neurotrophic Factor ; Immunohistochemistry ; Rabbits ; Stromal Cells ; enzymology ; Telomerase ; metabolism

Abstract: Objective　To develop a real-time fluorescent quantitative RT-PCR (qRT-PCR) method for qualitative and quantitative Chikungunya virus (CHIKV) analysis. Methods Based on the systematic analysis of the genomic sequences of Chikungunya and its related arboviruses, the specific nucleic acid sequences for Chikungunya virus were screened and identified, and then the primers and TaqMan probe were designed. Meanwhile, the human GAPDH gene was used as an internal reference. The reaction system for qRT-PCR was systematically optimized by L9(34) orthogonal design, and a rapid detection method for Chikungunya by qRT-PCR based on TaqMan probe methods was established. The sensitivity, specificity, reproducibility, and coverage of the established method were analyzed in detail. The standard curve was made, and the absolute quantitative method was established using the cloned nucleic acid fragments as positive samples. Results　A real-time fluorescent quantitative RT-PCR assay was developed for the qualitative and quantitative analysis of Chikungunya virus. The reaction system included Chikungunya virus and reference internal gene specific primers and probe, RT/Taq enzyme mixture, reaction buffer, and negative and positive reference. The established method obtained positive results with the ROSS strain of ECSA subtype, LR2006 strain of IOL branch, 181/25 strain of Asian type and Dongguan 2010 epidemic strains of Chikungunya virus, but there was no cross-reaction with other 18 arboviruses belonging to Flaviviruses, Alphaviruses and Bunyavirus. The minimum detection limit of the established method was 5.80 copies/mL, and a linear relationship was observed between the amount of input plasmid DNA and fluorescence signal value over a range of 5.80×102 copies/mL to 5.80×1010 copies/mL, and the correlation coefficient was 0.999 5. The qRT-PCR amplification efficiency was 91%, and the intra-assay variations and inter-assay variations were 0.01-0.07 and 0.03-0.11, respectively. Conclusions　The TaqMan qRT-PCR method developed in this study can qualitatively and quantitatively detect Chikungunya virus rapidly with specificity and sensitivity, providing a technical method for the prevention and control of this viral disease.

OBJECTIVETo explore cell death and apoptosis in rat hippocampal neurons at different time points after ischemia, hypoxia and reperfusion injury and to elucidate time window characteristics in ischemia neuronal injury.

METHODSHippocampal neurons were obtained from rat embryo and were cultured in vitro. The ischemia and reperfusion of cultured rat hippocampal neurons were simulated by oxygen-glucose deprivation (OGD) and recovery. OGD at different time points (0.25 h to 3.0 h) and then the same recovery (24 h) were prepared. Annexin V-PI staining and flow cytometry examined neuron death and apoptosis at different time after injury.

RESULTSAfter OGD and recovery, both necrosis and apoptosis were observed. At different times after OGD, there were statistically significant differences in neuron necrosis rate (P < 0.05), but not in apoptosis rate (P > 0.05). At recovery, survival rate of hippocampal neurons further decreased while apoptosis rate increased. Furthermore, apoptosis rates of different time differed greatly (P < 0.05). Apoptosis rate gradually increased with significant difference among those of different time points (P < 0.05). However, 2 h after ischemia, apoptosis rate decreased markedly.

CONCLUSIONSApoptosis is an important pathway of delayed neuron death. The therapeutic time window should be within 2 h after cerebral ischemia and hypoxia.

Animals ; Apoptosis ; physiology ; Brain Ischemia ; pathology ; Cell Death ; physiology ; Cell Hypoxia ; Cells, Cultured ; Disease Models, Animal ; Female ; Fetus ; cytology ; Flow Cytometry ; Hippocampus ; pathology ; Neurons ; pathology ; Pregnancy ; Pregnancy, Animal ; Probability ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology ; Sensitivity and Specificity ; Time Factors