Objective To establish a technical process for the separation of octopamine.MethodsTaking the absorptive capacity and elution efficiency of octopamine as indexes,the absorption characteristics and elution parameters of separation with macroporous resin were investigated.Results The static adsorption capacity of H103 type macroporous resin was 2.925mg g-1.The static elution ratio was 98.04%.Conclusion H103 type macroporous resin is effective to separate the octopamine and improve the flavour of fish sauce.

Objective To establish a HPLC method for the determination of octopamine in fish sauce.Methods A phenomenes luna C18 column was used.The mobile phase was 0.02?g?mL-1 citric acid-0.02?g?mL-1 sodium dihydrogen phosphate(7∶3,v/v) and detection wavelength was 274 nm.Results The linear range of octopamine was 104.0%,RSD=1.53%.The detection limit was 5.7ng?mL-1.Conclusion This method is simple,rapid and reliable.It could be used for the determination of octopamine and its related substances in fish sauce.The content of octopamine in Engraulis japonicus sauce is 1055?g?mL-1.

Objective To study the blood_flow_activating and blood_stasis_removi ng actions of Qushang Tablet (QT) and its safety. Methods Thirty rats were randomly allocated to Group A (normal control), Group B (models control) and G r oup C (treated with QT). Rat models with traumatic blood_stasis syndrome were in duc ed by the bruise of falling iron hammer. Effects of QT on blood rheology of rat model were studied. Acute and chronic toxic effect of QT was also studied. Res ults QT had obvious effect on blood rheology of rat model and could improve th e concentration, viscosity, aggregation and coagulability of blood. No acute and chronic toxic effect of QT was found. Conclusion QT exerts obvious blood_fl ow_activating and blood_stasis_removing actions and is safe to be applied in cli nic.

Objective:To observe the effects of acupuncture and moxibustion at Feishu(BL13)on inflammatory responses and intestinal short-chain fatty acids(SCFAs)in rats with asthma. Methods:Fifty-six Sprague-Dawley rats were randomly divided into a normal group(16 rats)and a modeling group(40 rats).Rats in the modeling group were subjected to establishing asthma models using ovalbumin(OVA).Model evaluation was conducted using 4 rats from each group.The remaining rats that successfully developed asthma were then randomly divided into a model group,an acupuncture group,and a moxibustion group,with 12 rats in each group.Rats in the acupuncture group received acupuncture treatments,and those in the moxibustion group received moxibustion treatments,both at Feishu(BL13)for 30 min.Following the treatments,the rats were exposed to atomization excitation with a 1%OVA solution for 20 min daily for 14 consecutive days.At the end of the experiment,inflammatory markers in the rats'peripheral blood were analyzed using a biochemical method.In addition,inflammatory cells in the bronchoalveolar lavage fluid(BALF)were counted using Wright-Giemsa staining.The lung tissue of rats was examined under a light microscope after staining with hematoxylin-eosin to observe morphological or pathological changes.Furthermore,real-time fluorescence quantitative polymerase chain reaction was utilized to measure the mRNA expression of inflammatory factors in the lung tissue.Lastly,the concentration of SCFAs in the rat's feces was determined using gas chromatography-hydrogen flame ionization. Results:The levels of eosinophils(Eos),neutrophils(Neu),and lymphocytes(Lym)in the peripheral blood,as well as Eos and Neu in the BALF,and the expression of interleukin(IL)-4,IL-5,IL-13,IL-33,and thymic stromal lymphopoietin(TSLP)mRNAs in the lung tissue were all found to be significantly increased(P<0.05 or P<0.01);the lung tissue structure displayed severe injuries;the levels of acetic acid,propionic acid,isobutyric acid,butyric acid,and valeric acid in the feces decreased significantly in the model group(P<0.05 or P<0.01).Compared with the model group,the peripheral blood levels of Eos,Neu,and Lym,as well as Eos in the BALF,and the mRNA expression levels of IL-4 and IL-5 in the lung tissue decreased significantly in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).This reduction was accompanied by alleviated pathological damage in the lung tissue.Additionally,there were significant increases in the levels of acetic acid,propionic acid,isobutyric acid,and butyric acid in the feces in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).In the acupuncture group,the expression levels of Lym in the BALF and IL-13 mRNA in the lung tissue decreased significantly(P<0.05 or P<0.01).In the moxibustion group,the mRNA expression levels of IL-33 and TSLP in the lung tissue also reduced significantly(P<0.05 or P<0.01).Furthermore,the level of valeric acid in the feces increased notably in the moxibustion group(P<0.01).Compared with the acupuncture group,it was found that the mRNA levels of IL-5 and IL-13 in the lung tissue,as well as the acetic acid level in the feces,were significantly higher in the moxibustion group(P<0.05 or P<0.01). Conclusion:Both acupuncture and moxibustion were effective in reducing abnormal inflammation and regulating intestinal SCFAs in asthma model rats.Acupuncture demonstrated superiority in inhibiting pro-inflammatory factors,particularly IL-5 and IL-13,while moxibustion exhibited better regulation on intestinal metabolites SCFAs,especially acetic acid.

BACKGROUND:It is reported that tailless-like protein (TLX) plays critical roles in the regulation of early developmental processes in vertebrates, and it plays a key role in stem cells proliferation and differentiation into neurons. OBJECTIVE: To construct recombinant plasmid pEGFPN1-TLX and study the transfection into dermal multipotential stem cells. DESIGN, TIME AND SETTING: Cytogene experiment was performed at the Department of Pathogen Biology, School of Basic Medical Science, the Third Military Medical University of Chinese PLA from March to December 2007. MATERIALS: An adult SD was obtained from the Experimental Animal Center of the Third Military Medical University of Chinese PLA; dermal moltipotential stem cells (DMSCs) were cultured by the Institute of Combined Injury of the Third Military Medical University of Chinese PLA; pEGFPN1 and DH5α was gifted by professor Xu.METHODS: Total RNA was extracted from rat brain tissue to amplify TLX-coded cDNA sequence using RT-PCR. T/A was cloned on pMD18-T vector and determined using BamHI and Hindlll. The products were positive recombinant plasmid pMD18-T-TLX segments, which were sub-cloned in pEGFPN1 to construct recombinant plasmid pEGFPN1-TLX. Finally, pEGFPN1-TLX was transfected into DMSCs.MAIN OUTCOME MEASURES: The fluorescence protein expression was observed under fluorescence microscope at 24 hours after transfection; TLX mRNA expression was detected using RT-PCR; neuronal differentiation was observed using immunohistochemical staining.RESULTS: TLX full length cDNA was successfully cloned into pEGFPN1, and pEGFPN1-TLX was successfully constructed by means of sequence analysis and enzyme cutting identification. As compared with non-transfected DMSCs, pEGFPN1-TLX transfected DMSCs were observed after 10 days, formed resistant clones after 15 days, and shown a green fluorescent protein expression. However, non-transfected DMSCs died at day 10. RT-PCR indicated that pEGFPN1-TLX transfected DMSCs could express TLX mRNA. At day 3 after induction, NF200 positive cells were increased, but glial fibrillary acidic protein positive cells were decreased after induction of pEGFPN1-TLX transfected DMSCs.CONCLUSION: TLX was successfully constructed and transfected into DMSCs. After transfection, neuronal differentiation of DMSCs was enhanced, and the differentiation to gliocytes was inhibited.

Objective To investigate the nutritional management level of Department of Clinical Nutrition in elderly inpatients through analyzing the actual hospital dietary intakes in elderly inpatients in Peking Union Medical College Hospital from 2013-2015.Methods Using continuous sampling, the nutrients contents and the eating rates of hospital meals ordered by the elderly inpatients on the 1st, 15th, 29th of every month from May 2013 to October 2015 were selected.The intake levels of energy, protein, calcium, vitamin A, vitamin B1 , and vitamin C were calculated and compared with the Chinese dietary reference intakes (DRIs).The nutritional in-take differences between diabetes diet and the basic hospital diet were also compared.Results A total of 90 days of hospital dietary data from 8 402 elderly inpatients were included in the study.The average eating rate was (50.1 ±4.2)%, which was lower than the general eating rate of the hospital [ (59.0 ±4.0)%, P<0.001]. Only protein intake from hospital diet reached the requirement target [male (103.1 ±47.3)%, femal (98.3 ± 33.8)%].Individual compliance rates were 63.2%and 59.8%, respectively, while the other kinds of nutrients were lower.The compliance rates of energy [ (73.3 ±26.3)%vs.(62.1 ±38.2)%, P<0.001] and ma-jor nutrient intake [ protein: ( 119.1 ±41.2 )% vs.( 93.3 ±65.1 )%, P <0.001; calcium: ( 55.5 ± 26.7)% vs.(34.3 ±34.2)%, P <0.001; vitamin A: (75.2 ±48.3)% vs.(57.4 ±97.1)%, P<0.001;vitamin B1:(76.4 ±38.2)%vs.(52.1 ±46.6)%, P<0.001;vitamin C:(92.2 ±51.4)%vs. (49.3 ±55.0)%, P<0.001) in diabetes diet group were significantly higher than those in the basic hospi-tal diet group.Conclusions The nutritional intakes from hospital diets may not meet the nutritional require-ments based on DRIs in elderly inpatients.Medical diet designed by physicians and dietitians should be ex-panded to improve the nutritional management level for these patients.

OBJECTIVE:To determinate the release rate and in vitro transdermal rate of asarinin in Cancer pain cataplasm. METHODS:Using homemade devices and modified France diffusion,isolated skin of rats as barrier,normal saline as solvent,the content of asarinin was determined by HPLC. Release rate of Cancer pain cataplasm within 20,50,80 and 120 min and transder-mal amount within 2,4,8,12,24 h were investigated,and accumulative release rate and accumulative transdermal rate were cal-culated. RESULTS:Accumulative release rate by 120 min of asarinin in Cancer pain cataplasm was 73.01%;24 h in vitro transder-mal rate was 26.01%,and transdermal kinetics equation of asarinin was Q=5.717 7t1/2-0.385 4(r=0.979). CONCLUSIONS:Cancer pain cataplasm has good release and transdermal performance. Its transdermal kinetics is in line with Higuchi equation.

BACKGROUND:Bone marrow mesenchyme stem cells are important non-hematopoietic stem cells in the bone marrow, which can stimulate angiogenesis. While, Slit can also stimulate angiogenesis, as many studies have proved. OBJECTIVE:To review the biological functions, clinical application and effects of bone marrow mesenchymal stem cells and Slit2 on promoting angiogenesis. METHODS:A computer-based online research of CNKI and PubMed databases was performed to col ect articles published between 1980 and 2014 with the keywords“MSCs”and“Slit2”in Chinese and English. There were 436 articles after the initial survey. Final y, 65 articles were included according inclusion and exclusion criteria. RESULTS AND CONCLUSION:Both bone marrow mesenchymal stem cells and Slit2 play an important role in promoting angiogenesis, but the relevance of bone marrow mesenchymal stem cells and Slit2 is stil controversial. If assuming that bone marrow mesenchymal stem cells secrete Slit2, more researches should be done to reveal whether bone marrow mesenchymal stem cells promoting angiogenesis is relevant to Slit2 and through which signaling pathway Slit2/Robo functions to adjust bone marrow mesenchymal stem cells thus to promote angiogenesis. If relevant, the transplantation of the Slit2 and bone marrow mesenchymal stem cells wil be a promising treatment of cerebral infarction and other central nervous injuries.

The primary action of corticotropin releasing hormone (CRH) is stimulation of the synthesis and release of adrenocorticotropic hormone and β-endorphin from the pituitary in response to stress. In addition, a number of studies indicate that CRH exerts other physiological actions within the central nervous system which are independent of the pituitary. These include increased body temperature and thermogenesis. However, the intracellular mechanism responsible for pyrogenic action of CRH is still unclear. The purpose of these studies was to determine whether or not cAMP was involved in the pyrogenic action of CRH in the rat. Intracerebroventricular (icv) microinjection of CRH (2.5 μg, 5.0 μg, 10 μg) caused increases in colonic temperature and hypothalamus cAMP level in conscious rats. The pyrogenic effects of CRH were abolished or markedly inhibited by prior injection (icv) of an adenylate cyclase inhibitor, 2，，3，-dideoxyadenosine (DDA, 30 μg) or an inhibitor of cAMP-dependent protein kinase, adenosine-3，，5，-(cyclic) monophosphorothionate (Rp-cAMPs, 15 μg). This is the first report demonstrating the pyrogenic effcet of centrally administration of CRH on the rat via the cAMP-mediated protein kinase signal transduction pathway.

Objective To observe the effect of low intensity pulsed ultrasound (LIPUS) on chondrocytes co-cultured with infrapatellar fat pads.Methods Twenty-four infrapatellar fat pads and cartilages from female knee trauma patients aged between 25 and 35 were cut and stained using hematoxylin-eosin staining.Chondrocytes were isolated from part of the integrated surface of the cartilages to be cultured in vitro.They were then randomly divided into a normal chondrocyte group (the control group),a normal chondrocyte+FCM (fat conditioned medium) group (the model group),a normal chondrocyte+ FCM + LIP US group (the treatment group),and a normal chondrocyte+ FCM + LIPUS + GRGDSP (an integrin inhibitor) group (the inhibited group).The treatment group and inhibited group received LIPUS at 40 mW/cm2 for 20 min once a day,while the other groups received sham LIPUS treatment.Five days later,the cells were collected and western blotting was used to examine the expression of type Ⅱ collagen (COL2),aggrecan (Acan),matrixmetalloproteinase (MMP)-13,integrin β1,phosphorylation-focal adhesion kinase (p-FAK) and phosphorylation p38 (p-p38).Results Western blotting showed that compared with the control group,the expression of COL2 and Acan was significantly lower in the model group,but that of MMP-13,integrin β1,p-FAK and p-p38 was significantly higher.As compared with the model group,the expression of COL2,Acan,integrin β1 and p-FAK was significantly higher,and that of MMP-13 and p-p38 was significantly lower in the treatment group.The expression of COL2,Acan,MMP-13,integrin β1,p-FAK and p-p38 showed no significant difference between the inhibited and model groups,but that of COL2,Acan,MMP-13,integrin β1,p-FAK and p-p38 was significantly different between the control and treatment groups.Conclusions LIPUS provides a protective effect on chondrocytes through inhibiting the expression of MMP-13 induced by adipokines and the degradation of COL2 and Acan through activating the integrin-FAK-p38 MAPK pathway.