Objective To observe whether endomorphin-2-immunoreactive(EM2-ir) neurons in the trigeminal ganglion(TG) project to the medullary dorsal horn(MDH) in the rat. Methods Fluoro-gold(FG) retrograde tracing and immunofluorescent histochemical staining for EM2 were applied. Results After FG was injected into the MDH,many FG labeled neurons with small,medium and large diameters were observed in the TG.There were also aboundant EM2-ir neurons with different diameters in the TG.The majority of FG labeled neurons in the TG showed EM2-immunoreactivities.Almost all of the EM2/FG double-labeled neurons were medium and small in size.Conclusion EM2-ir neurons in the TG project to the MDH in the rat.

Objective To isolate protoplasts from tube plant leaves of Rhodiola sachalinensis and regenerate plantlets after protoplast culture.Methods Preculture treatment and size of explants,enzyme concentration,mannitol concentration in enzyme mixture related with protoplasts isolation were studied to determine the superior optimized conditions.Results Explants could be used to isolate protoplast without dark preculture,and leaf length should be longer than 1.5 cm.The best incubating enzyme solution contains 1.0% cellulase Onzuka R-10,0.5% Macerozyme R-10,10 mmol/L CaCl2?2H2O,0.1% MES,0.7 mmol/L KH2PO4,and 0.5 mol/L mannitol.The enzyme and explants mixture were shaken for 4 h at 25 ℃.The protoplasts yield and viability were 39.43?106/g fresh weight and 78.6%,respectively.Purified protoplasts were cultured in medium 1/2 MS+1 mg/L 2,4-D+0.5 mg/L ZT+0.5 mol/L mannitol +500 mg/L hydrolysis of casein initially with shallow liquid layers,and calli formed within 40 d.After calli were transfered to MS+1 mg/L 6-BA+0.1 mg/L NAA,adventitious buds were induced from calli.Shoots longer than 2 cm rooted within 30 d when they were transfered to 1/2 MS medium.Conclusion The study provides the scientific base for protoplast fusion in polyploidy breeding of R.sachalinensis.

Objective To investigate the characteristics of intestinal flora in infants with different feeding patterns.Methods Sixty-two cases of health infants(30-120 d)were divided into 4 groups according to their feeding patterns:breast feeding,imported powder milk feeding,domestic powder milk feeding and mixed feeding.Samples of their fresh feces in each group were collected and divided into sections equally:the bifidobacteria were isolated in anaerobic box and the number was counted for one section;for the other section,total DNA of intestinal flora was extracted and enterobacterial repetitive intergenic consensus (ERIC) fingerprints were amplified with the method of ERIC-PCR.After that,the specific bands observed in different groups were cloned and sequenced and alignmented.Results The colonies of bifidobacteria were more in breast feeding and mixed feeding groups[(9.10 ± 1.33) cfu/g;(8.62 ± 1.35) cfu/g]than those in domestic powder milk feeding and imported powder milk feeding groups[(7.62 ± 1.22) cfu/g;(7.32 ± 0.80) cfu/g,t =3.23,P ＜ 0.05];while there was no significant difference between breast feeding and mixed feeding groups,and between 2 powder milk feeding groups.Two specific bands were found from the ERIC fingerprints (A:1 100 bp mainly in breast feeding,domestic powder milk feeding and mixed feeding groups;B:1 000 bp mainly in imported powder milk feeding group).Sequencing and analysis of Basic Local Alignment Search Tool showed that homologous bacteria of A and B fragments were bifidobacterium longum.The encoding protein of A fragments might be related to the enzymes of carbohydrate metabolism,and B fragments were related to the enzymes of protein metabolism.Conclusions The colonies of bifidobacteria in intestinal tract are more in breast feeding and mixed feeding infants than those in formula feeding groups.The distribution of intestinal flora in domestic powder milk feeding infants is more similar to that of the breast feeding infants.

Objective To investigate the effect of mixed application of lecture-based learning and flipped classroom and problem-based learning in microbiology teaching. Methods The five-year program students of Grade 2013 and Grade 2014 were randomly divided into two groups, and the experimental group (48 students in the teaching reform class) carried out the blended teaching, while the contrast group (48 students of parallel class) carried on the traditional teaching. After the lecture, the teaching effect of the hybrid teaching method and traditional teaching method was analyzed after the theoretical and the experi-mental operation test were adopted, and the four part questionnaire surveys including the integrated use of knowledge, active classroom atmosphere, innovation ability and teaching satisfaction were proceeded in each group. The data of each group was analyzed by t test analysis with SPSS 19.0 respectively. Results The theory test scores of experimental group was (90.16±3.14), which was higher than the control group (82.33± 4.21). The difference between them was very significant (P=0.000). Survey results showed that the integrated use of knowledge, active classroom atmosphere, innovation ability and teaching satisfaction were higher than traditional group. The difference was statistically considered significant (P<0.01). Conclusion Hybrid teaching method is very good for training and development of students' comprehensive quality and ability. It has important significance in improving the students' score.

The present study was designed to examine the morphological pattern of primary afferent projections into the spinal dorsal horn by labeling the lumbar dorsal root ganglia with carbocyanine fluorescent dye DiI in mouse embryos and neonatal pups aged embryonic day 12 to postnatal day 3 (E12-P3). Primary afferent fibers projected into dorsal funiculus at E13, but did not penetrated into gray matter of dorsal horn until E15. The afferent projections became dense and entered the spinal gray matter more deeply at E16 and E17. By E18 the intensity of primary afferent in the deep part of the dorsal horn increased and their branching patterns became more complicated. Some of these primary fibers were also observed to ramify extensively in the superficial laminae. The projection pattern of primary afferent remained unchanged after birth, but the intensity of afferent terminals increased in the superficial laminae. In addition, afferent fiber collaterals that projected into the contralateral dorsal horn were also observed. They were first examined at E16 and mainly originated from the medial and deep part of the dorsal horn. Around birth, the contralateral projections were also found to originate from the lateral part of dorsal horn. Our results indicate that laminar organization of primary afferents in the spinal dorsal horn is established during the late embryonic and early postnatal stages.This organization then undergoes further refinement to match the pattern seen in the adult.

The paper analyzes the application requirements of ECG examination in the wards of Southwest Hospital and describes the application of the remote ECG system in the wards.It introduces the system's composition,functional modules,business process,key technology and application effects.The application of the remote ECG system can reduce the waiting time of patients and realize the ECG examination at the bedside.Thus,it is well received by patients and clinical departments.

Objective:To clone Fab genes of anti-p185 monoclonal antibody 5E12 and express it in E.coli.Methods:Fd and ? genes were cloned by RT-PCR, inserted into Fab expression vector and expressed in E.coli. The N-terminal sequences of V regions was resumed by PCR mediated mutagenesis. The antigen-binding activity of the Fab were tested by ELISA and immunohistochemistry.Results:Fd and ? genes were cloned and expressed in E.coli. But the bacterially expressed Fab fragments showed no antigen binding activity. After the N-terminal sequences of V regions was corrected to original sequences, the Fab expressed in bacterial was able to target HER2/neu-expressing cells(NIH3T3/erbB-2 cells). Correction of Fd N-terminal sequences could partially resume the antigen binding activity. But correction of ? chain N terminal sequences was shown no expected result.Conclusion:Successful in constructing and expressing anti-p185 Fab, which will benefit the construction of other engineering antibody and humanization of murine anti-p185 McAb. We also found that the V region N terminal changes introduced with PCR primers may affect antigen binding activity seriously, to which more attention should be paid during antibody engineering.

Objective:To investigate the antitumor effect of TKD-activated NK cells on tumor growth inhibition of human pancreatic carcinoma in nude mice .Methods:CD3-CD56+NK cells were obtained from human peripheral blood mononuclear ( PBMC) in stem cell growth medium SCGM , 2 μg/ml TKD was added to the medium on day 10.The activating receptor CD 94/NKG2C expression levels on NK cells was detected with FAC after 4 days.The cytolytic activity of TKD-activated NK cells against human pancreatic carcinoma subline with HSP 70 expression on their cell surface was analyzed by MTT assay .Established a new model of orthotopic-transplantation tumor of human pancreas .NK cells were injected i.v.into the tail vein of tumor-bearing mice on day 15,the antitumor activity of the NK were evaluated .The capacity to infiltrate Colo 357 tumors in SCID/beige mice was detected with Immunohis-tochemistry.Results:Flow cytometry analysis showed that CD3-CD56+NK cells could expanded in SCGM medium ,and the average percentage of NK cell was (87.50 ±1.35 )%.CD94 expression levels on NK cells increased obviously ,the mean fluorescence intensity of CD94 was(220.56±1.82),compared to (68.72±1.85)of control group cell.The cytolytic activity against HSP70 membrane-positive pancreatic carcinoma sublines Colo 357 cells was high and there was significantly statistical difference between TKD-activated NK cells and unactivated NK cells.The cytolytic activity was(68.72±2.55)%when ratio of effector cells and target cell was 40:1.TKD-activated NK cells had a stronger suppressive effect on tumor growth in BALB /c nude mice bearing Colo 357 cells in vivo ,Median inhibitory rates was ( 61.3 ±1.5 )% .There was significant statistical difference compare to control group ( P <0.01 ) .The result of Immunohistochemistry indicated that predominantly NK cells induced with TKD had the capacity to infiltrate Colo 357 tumors in SCID/beige mice.Conclusion: TKD-activated NK cells are highly efficient cytolytic effector cells which have a stronger significant suppression against pancreatic carcinoma growth in vivo .

Objective To investigate the synaptic connections between endomorphin-immunoreactive(ir) terminals and ? opioid receptor-ir neurons in the superficial layers(laminae Ⅰ and Ⅱ) of the rat spinal dorsal horn. Methods The double-labeling immunohistochemical electron microscopic technique,i.e.pre-embedding immunohistochemical detection of endomorphin combin with pre-embedding immuno-gold particles labeling technique to identify ? opioid receptor-immunoreactivity,was used in the present study. Results ? opioid receptor-ir neuronal cell bodies,fibers as well as axon terminals were observed in laminae Ⅰ and Ⅱ of the spinal dorsal horn,meanwhile dense endomorphin-ir axon terminals were located mainly within the same region.Under electron microscope,the synaptic connections formed by endomorphin-ir axon terminals labeled with diaminobenzidine(DAB) reaction products and ? opioid receptor-ir neuronal cell bodies and dendritic profiles labeled with immuno-gold particles were observed.The principle synaptic type was asymmetric synapse.There were more axon-dendritic synapses than axon-somatic ones as encountered in the present study.Conclusion The distribution patterns of endomorphin-ir and ? opioid receptor-ir structures match with each other in the superficial laminae of the spinal dorsal horn,that provides morphological base for the antinociceptive effects of endomorphin and ? opioid receptor in the processing of nociceptive information transmission.