BACKGROUND:Bone marrow mesenchymal stem cells (BMSCs) of mice are different from human and rats.The difficulties of culture,sustaining impermanency activity of BMSCs after passage and short-term effect of stem cells tracking limited the study of mice.OBJECTIVE:To obverse modified isolated culture method of BMSCs of mice and the feasibility of long-term fluorescent labeling stem cells.DESIGN,TIME AND SETTING:The experiments were conducted at the Affiliated Hospital of Academy of Military Medicine Science from June to December in 2008.MATERIALS:C57BL/6 mice,males and females,4-6 weeks of age,mean weighing 20 g,were used.METHODS:Stem cell culture fluid serum and liquid change manner were optimized using adherent screening and Percoll separation method.Rat BMSCs were incubated.In accordance with previous experiences of MSCs between human and macaque,Hyclonehigh-grade fetal bovine serum was selected for mouse MSC incubation.Serum was 10% of the medium.Bone marrow cells were washed out using LG-DMEM to filter bone dregs and small muscle blocks.Subsequently,samples were added on the Percoll separating medium at relative density of 1.082,and then incubated in 75 cm~2 culture flask at the concentration of 1.5×10~6/cm~2.BMSCs at the second passage were labeled with CM-Dil.MAIN OUTCOME MEASURES:Morphological changes in primary cultured and subcultured mouse BMSCs were measured.BMSC surface antigen of the second passage of mice (CD105,CD44,CD25,CD34) were determined using flow cytometry.The modified method was assessed to harvest the purity of stem cells.Activity of mouse BMSCs was identified using adipogenic and osteoplastic differentiation.The strength of fluorescent cells following multiple passage was observed.RESULTS:①The attached cells were observed 48 hours after primary cells culture and changed shuttles,triangles and flats at 7 days after culture.The figure become bunches and radial pattam at 3 passages.②MSCs highly expressed CD105,CD44 phenotypes and seldom expressed CD34 and CD45.③Spindle shape of cells gradually disappeared,with increased cell body.Some cells grew in cluster.MSCs changed figures to multilayer knots 10 days after inducing osteoplastic differentiation.MSCs became roundness and appeared fat drops in cells 9 days after inducing adipogenic differentiation.Red fat particles were shown following oil O staining.④The labeling cells gave out oranges light,and marked rates were over 80% in MSCs under the fluorescent microscope.The labeling cells were over 47% in 4 passages MSCs using flow cytometry.CONCLUSION:The modified method gained high-dosage cells in shorten culture time at passage 2 and made CM-Dil long lime labeling cells,which made more convenient for MSCs experiment on mice and stem cells tracer experiment in vivo.

A rapid and sensitive analytical method was developed for the simultaneous determination of fumonisins B1 and B2 in traditional Chinese medicines by HPLC-MS/MS. The detection limits for fumonisins B1 and B2 were 0.25 ng x mL(-1) corresponding to 2 microg x kg(-1) in samples. Recoveries from different samples spiked with fumonisins B1 and B2 at levels ranging from 0.2 to 3 mg x kg(-1) were 84.0%-96.1% and 86.3%-99.3%, respectively. Among the total of 34 samples purchased from local markets, ten samples of which were visibly moldy samples due to inappropriate storage, and 24 were normal samples. The results showed that 6 of the visibly moldy samples and 5 of the normal samples were contaminated with total fumonisins at levels ranging 82.4-2349 microg x kg(-1) and 102-729 microg x kg(-1), respectively.

Objective To explore cancer factor exhausted during chemotherapy intermittent for drinking, food and oxygenic sport interference to patients suffered from ovarium cancer. Method 92 patients suffered from ovarium cancer with drinking, food and oxygenic sport interference and taking self-comparison method. Simple tired Assessment Form, European cancer treatment and research origination questionnaire (EORTC-QLQ-C30) are as-sessed for the patients suffered from cancer tied and life quality situation before interference and after interference of 30 d so as to observe the tired relieving degree and life quality of the patients before and after interference. Results Using nursing interference and then the patients relive their tiredness greatly to have statistical significance (P <0.01). The comparison before and after interference and the life quality has statistic significance(P <0.01). Con-clusion The drinking, food and oxygenic sport interference can relieve the fired degree of the patients suffered from ovarium cancer effectively to improve the life quality of the patients.

Objective To know the influence of psychological nursing intervention for quality of life in patients with ovary cancer. Methods Divided 300 patients with ovary cancer into the intervention group and the control group randomly, there were 150 cases in the each group. Routine nursing cares was ued in the control group, the psychological nursing intervention was used in the intervention group in addi-tion. Compared the quality of life between the two groups by interviewed questionnair. Results After the nursing intervention, the indexes which can indicated the quality of life in the intervention group were bettez than those of in the control group significantly. Conclusions Psychological nursing intervention can ef-fective promote the quality of life of patients with ovary cancer.

Objective To evaluate the role of alpha4 beta2 neuronal nicotinic acetylcholine receptor in the inhibition of synaptic long-term potentiation (LTP) by isoflurane in the CA1 area of rat hippocampal slices.Methods Hippocampal slices (400 μm thick) were prepared from the brains of adult male SD rats, 2 months old, weighing 200-250 g, anesthetized with ether and decapitated. The slices were incubated in artificial cerebrospinal fluid (aCSF) at room temperature for at least 2 h before use. Seventy slices were randomly divided into 10 groups ( n = 7 each): Ⅰ LTP group in which the slices were perfused with aCSF; Ⅱ , Ⅲ and Ⅳ group in which the slices were perfused with aCSF containing isoflurane 0.125, 0.25 and 0.5 mmol/L respectively (group Ⅰ1-3 );Ⅴ and Ⅵ group in which the slices were perfused with aCSF containing epibatidine 0.1 and 1.0 μmol/L respectively (group E1.2 ); Ⅶ group epibatidine 0.1 μmol/L + isoflurane 0.25 mmol/L (group E1 + I2 ); Ⅷgroup epibatidine 1.0 μmol/L + isoflurane 0.25 mmol/L (group E2 + I2); Ⅸ group DHβE 0.1 μmol/L (group D); Ⅹ group DHβE 0.1 μmol/L + isoflurane 0.125 mmol/L (group D + I1 ). Population spikes (PS) were recorded for at least 30 min before LTP in each group. For LTP induction, high-frequency stimulation (HFS) was applied to the Schaffer collateral-commissural pathway of hippocampus and maintained for 15 min using a stimulating electrode.The changes in PS amplitude were analyzed at 5, 10, 15, 20, 25, 30, 40, 50 and 60 min after HFS in each group. Results Compared with group LTP, the PS amplitude was significantly decreased after HFS in group I1 ,I2, I3 , D, D + I1 and E1 + I2 ( P ＜ 0.05), while increased after HFS in group E1 .2 ( P ＜ 0.05 ), but no significant change was found after HFS in group E2 + I2 ( P ＞ 0.05). The PS amplitude was significantly decreased after HFS in group D + I1 compared with group I1 (P ＜ 0.05). The PS amplitude was significantly increased after HFS in group E1 + I2 and F2 + I2 compared with group I2 ( P ＜ 0.01 ). Conclusion Isoflurane inhibits LTP induction via inhibiting the activation of alpha4 beta2 nicotinic acetylcholine receptor in rat hippocampus.

Objective To investigate the clinical efficacy of combined detection of VCA-IgA and Rta-IgG in the diagnosis of nasopharyngeal carcinoma.Methods From May 2013 to November 2014, 3 913 serum samples(male 2 367,female 1 546) from healthy people who had health examination in our medical center were collected and 169 serum samples(male 118,female 51) were collected from the patients who were diagnosed as nasopharyngeal carcinoma by pathological biopsy.Serum samples in two groups were detected by EBV RTA-IgG, VCA-IgA assay ( ELISA ) respectively.SPSS17.0 statistical software and receiver operating characteristic curve ( ROC) were applied to data analysis.Results The Rta-IgG positive rates of EB virus were 93.5%in NPC group (158/169) and 2.4%(93/3 913) in healthy group;while the VCA-IgA positive rates were 79.3%in NPC group ( 134/169 ) and 8.9% ( 349/3 913 ) in healthy group. The sensitivity(χ2 =14.49,P<0.05) and specificity(χ2 =157.15,P<0.05) of Rta-IgG in the diagnosis of nasopharyngeal carcinoma were significantly better than that of VCA-IgA. Using VCA-IgA/Rta-IgG combined detection analysis, not only failed to effectively improve the diagnosis of nasopharyngeal cancer, but to reduce the detection sensitivity to 72.8%( 123/169 ) , compared with Rta-IgG detection only. Conclusions Rta-IgG is significantly better than that of VCA-IgA.There was no significant improvement in the clinical diagnostic efficacy of nasopharyngeal carcinoma using VCA-IgA/Rta-IgG combined detection mode.

Objective Compared with haloperidol treatment,naloxone and naloxone combined haloperidol treatments were assessed in their efficacy and safety for excited type delirium in elderly.Methods The elderly patients with delirium were divided into haloperidol treatment (H),naloxone treatment (N) and combined treatment (C) in a prospective randomized controlled design.Delirium score scale (DSS) was used before and after treatments respectively.Clinical global impression scale-severity of illness (CGI-SI) score was evaluated daily to assess the onset time and improvement of delirium.Agitation-calmness evaluation scale (ACES) observed calmness effect in agitated patients before and after every medication intervention.Treatment emergent symptom scale (TESS) assessed side effects of all medications.Results The duration of three groups(H(4.0 ±2.9)d,N(4.2± 3.5) d,C (3.2 ± 3.2) d) had no significant difference (P ＞ 0.05) by multiple comparison.Compared the onset time of three groups (H (2.4 ± 1.6) d,N (2.4 ± 1.4) d,C (1.3 ± 0.9) d),the combined group was faster than the other two groups (all P ＜ 0.05),no significant difference between the two groups(P ＞ 0.05).DSS scores had no significant differences (P＞ 0.05) in three groups before treatment,so did CGI-SI scores.In the end,DSS scores were(H:18.8 ± 11.5,N:27.7 ± 7.2,C:29.5 ± 5.6) respectively.Statistically naloxone group and combined group with no significant difference (P ＞ 0.05),were better than the haloperidol group (all P ＜ 0.01) in recovery.At the same time CGI-SI scores were (H:3.3 ± 1.5,N:2.5 ± 1.5,C:1.8 ± 0.9) respectively.Statistically combined group was better than the haloperidol group (P ＜ 0.01),and no significant difference with naloxone group (P ＞ 0.05).Three groups had no significant difference in ACES scores (P ＞ 0.05) before and after medication interventions(H:5.9 ± 1.6,N:6.2 ± 1.8,C:6.4 ± 1.6) (P ＞ 0.05).Haloperidol group had 4 cases of extrapyramidal symptom evaluated by TESS and combined group had 2 cases.Two groups had no significant difference (Chisquare test P ＞ 0.05) in extrapyramidal symptom incidence.The naloxone group showed no side effects.Conclusion Naloxone combined haloperidol is slight better than naloxone,more than haloperidol in terms of efficacy.Haloperidol equally with combined medication showed more side effects than naloxone.

An 81-year-old male presented with an 8-year history of recurrent ulcer on the left dorsal foot which gradually spread to involve both lower limbs. Physical examination revealed no abnormality of any organ systems and no palpable superficial lymph nodes. Skin examination showed erythematous swelling of the left dorsal foot with an ulcer sized 7 cm × 10 cm on the surface. Tendon was visible at the base of the ulcer, and the ulcer margin was elevated giving a dyke-like appearance. The perilesional skin was purple-brown. There were several millet-like papuloid lesions circularly arranged at the inner side of the right foot as well as dark erythematous or brown nodules and pigmented patches with tenderness on both lower limbs. Histopathology of the ulcer of the left dorsal foot and papuloid lesions on the right foot revealed a visible epidermotropic infiltrate in the epidermis as well as an infiltration throughout the entire dermis with medium-sized atypical lymphoid cells with obvious mitoses. Immunohistochemical examination showed the coexpression of both T cell markers (including CD3, CD45RO, CD43) and B cell marker (CD20), with scatted positive staining for PAX-5and negative staining for CD79α or CD1 9. PCR confirmed the rearrangement of T cell receptor (TCR)-γgene. A diagnosis of peripheral T cell lymphoma unspecified was made in view of the rearrangement of TCR-γgene and above findings. The patient was treated with the following modified CHOP regimen: intravenous cyclophosphamide 0.8 g, leurocristine 2 mg and epirubicin hydrochloride 60 mg, as well as oral prednisone 15 mg twice daily for 5 days every 3 weeks (one treatment session). After 3 treatment sessions, the lesions improved markedly.

Faciliysyion techniques were used to help improve the extremity motor functions of 43 elderly and 79 middle aged or youthful stroke patients. The Fugl-Meyer scale was employed for thc assessment of the functional status. The results showed that the motor function of upper extremty of thc elderly patients was not improved. but the motor function of lower extremity was improved significantly. The motor functions of upper and lower extremities were all improved significantly in the middle-aged or youthful patients. In early rehabilitation group the Increased motor function marks of the upper extremity of the elderly stroke patients were less than that of the middle-aged or youthful stroke patients. the increased motor function marks of the lower extremity of The elderly stroke patients were less significantly than that of the middle-aged or youthful stroke patients also. In late rehahilitation group the increased motor function marks of the lower extremity in elderly stroke patients were more than that in the middle-aged or youthful stroke patients.

Objective To study the bacteriological characteristics and the pathogenesis of Staphylococcus aureus (S. aureus) on eczema and atopic dermatitis (AD). Methods A multi-center randomized, double blind bacteriological study on the lesions and non-lesional skin of patients with eczema (207) and AD (119) were carried out. The antibiotic sensitivity and the bacteriophage typing were performed on all the S. aureus isolated from the patients. Results There were statistical differences in the positive rate of the culture, the ratio and the colonization of S. aureus between the lesion and the non-lesional skin in eczema (P