This paper employed UPLC-Electrospray Ionization /Quadrupole-Time of Flight-Mass /Mass Spectrometry( UPLC-ESI/Q-TOF-MS/MS) to analyze the chemical constituents in the stems of Nelumbo nucifera. The stems of N. nucifera were extracted with 75% methanol, and we applied an Agilent Zorbax SB-Aq column (2.1 mm x 100 mm, 1.8 μm) to UPLC analysis with water methanol-water( containing 0.05% formic acid) in gradient as mobile phase. The eluates were then detected by ESI-Q-TOF-MS/MS. Results indicated that 22 benzylisoquinoline alkaloids were indendified. Among them, one alkaloid may be a new compound and a component was found in the Lotus for the first time. We fully identify the composition of the Lotus stems for the first time, Which could provides theoretical foundation for further study and utilization of the medicinal resources.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Nelumbo ; chemistry ; Plant Stems ; chemistry ; Tandem Mass Spectrometry ; methods

To clarify the origin and development of the traditional Chinese medicine "Duhuo" and "Qianghuo" with medicinal literatures. Medical literatures of past dynasties were analysed and combined with the modern material. The "Duhuo" in Herbal writing Shen Nong Ben Cao Jing include traditional Chinese medicine "Duhuo" and "Qianghuo", "Qianghuo" was separated from "Duhuo" due to the distinguish of clinical application. The origin of "Qianghuo" is Notopterygium incisum and N. forbesii, However, The origin of "Duhuo" is very complex, Angelica pubescens f. biserrata as authentic "Duhuo" was used from Song Dynasty. "Qianghuo" was originated from "Duhuo".
Angelica ; chemistry ; growth & development ; Apiaceae ; chemistry ; growth & development ; China ; Drugs, Chinese Herbal ; isolation & purification ; Geography ; Humans ; Medicine, Chinese Traditional ; Research ; Species Specificity

BACKGROUND:Periosteum is considered as a source of seed cels for cel therapydue toits biological features. OBJECTIVE:To seek the optimal way to isolate and culture rabbit periosteal cels and identify their biological features. METHODS:Rabbit periosteum on facies medialis tibiae was taken out under aseptic conditions. Periosteal cels isolated through the digestion of type II colagenase with the explants culture method were cultured in DMEM/F12 complete medium. Cel ultrastructure was observedunderan inverted microscope. Periosteal cel proliferation was determined bycel counting kit-8assay. Cel surface antigensCD90 and CD105 were determined using flow cytometry. Osteogenic andlipogenic induction mediums were applied to induce periosteal cels to differentiate into osteocytes and adipocytes, respectively. After 2 weeks of induction, cels were harvestedfor alizarin red staining and oil red O staining to assay the calciumnodules and lipid droplet. RESULTS AND CONCLUSION:The digestion of type II colagenase with the explants culture method shortened the period of primary cels culture and enhanced the survival rate, which causedhigher purity and stronger reproductive activity of harvested periosteal cels. Primary cultured periosteal cels grew in form of spindle spiral or paralel. Alizarin red andOil red O staining verified the multi-directional differentiation potentiality of periosteal cels. These findings suggest that the periosteal cels with high purity,strong reproductive activity,andmulti-directional differentiation potentialitycanbe harvested in short time using digestion of type II colagenase with the explants culture method.

AIM: To observe the effect of microRNA-16 (miR-16) on the megakaryocytic differentiation of K562 cells, and to explore the potential mechanism.METHODS:miR-16 was over-expressed or silenced by transfection with miR-16 mimics or inhibitor in K562 cells.The level of miR-16 was detected by real-time PCR.The expression of CD41, CD42b and CD61, as megakaryocytic differentiation markers, was detected by flow cytometry.The effect of miR-16 on the expression of myeloblastosis oncogene ( MYB) was measured by Western blotting, and flow cytometry was performed to confirm whether the effect of miR-16 on expression of CD41, CD42b and CD61 was mediated by MYB.RESULTS:Transfection with miR-16 mimics dramatically elevated the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells.Transfection with miR-16 inhibitor decreased the level of miR-16 and the expression of CD41, CD42b and CD61 in the K562 cells (P<0.05).The expression of MYB was regulated by miR-16, and MYB silencing reversed the regulation of CD41, CD42b and CD61 induced by miR-16.CONCLUSION:miR-16 regulates the megakaryocytic dif-ferentiation of K562 cells by targeting MYB.

Objective To study the effect of Punica granatum( pomegranate) see d oil( PSO) on proliferation and apop-tosis behaviors of breast cancer cells.Methods Fatty acid composition was detected by gas chromatography,breast cancer cells, MCF-7 and MDA-MB-231 were treated with PSO, cell proliferation was observed by MMT, cell apoptosis was analyzed by flow cytometry,and expression levels of proliferation and apoptosis-related proteins were detected by Western blot.Results Punicic acid (PA) was the major fatty acid in PSO(74.41%).PSO could inhibit the proliferation while in-ducing apoptosis in both cell lines in a dose-and time-dependent manner, significantly decrease the expression level of Cox-2 and Bcl-2, increase the expression level of Bax and caspase-3 (cleaved),remarkably upregulate the expression of P53 in MCF-7, and downregulate p53 expression in MDA-MB-231.Conclusion PA may be one of the functional ingredients of PSO which can inhibit proliferation and induce apoptosis in breast cancer cells.These effects are probably mediated by regu-lating the expression of Cox-2, Bcl-2, Bax, caspase-3 (cleaved) and p53.

Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.

Lonicerae Japonicae Flos was one of the most widely used traditional Chinese medicine for its special biological activities. The content of luteoloside, one of its major compounds, was an important standard for the quantity control of Lonicerae Japonicae Flos. The major method used for the detection of luteoloside was instrumental analysis. Compared with the ELISA method, instrumental analysis was time-consuming, complex pretreatment and low-throughout. Thus, it was significantly important to develop an enzyme-linked immunosorbent assay (ELISA) for luteoloside analysis. Here, the conjugates of luteoloside-bovine (LG-BSA) and luteoloside-ovalbumin (LG-OVA) were produced as the immunogen and coating antigen by the carbodiimide ( CDI) method, respectively. The conjugation ratio of carrier protein and the hapten in the conjugate were determined by UV-Vis spectrophotometry (UV). LG-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The titer and specificity of antiserum were detected by ELISA. The conjugation ratio of hapten and carries protein were 3. 7: 1 (LG-BSA) and 1. 0: 1 (LG-OVA). The antiserum titer was higher than 2 000 with the linear range of 18.4-4 852.4 μg x L(-1), R2 = 0.988 4 and IC50 = 298.7 μg x L(-1). The result showed that the conjugate antigen LG-BSA was synthesized successfully and the mice can produce specific antiserum injected with artificial antigen.
Animals ; Antibodies ; analysis ; immunology ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunization ; Lonicera ; chemistry ; immunology ; Mice ; Mice, Inbred BALB C

BACKGROUND: Upper eyelid flap grafting-related vessels such as superficial temporal artery, supratrochlear artery, supraorbital artery trunk are reported. Upper eyelid artery dissection is becoming more and more important for the surgery on the eyelid, but there is a lack of anatomical analysis of upper eyelid artery. OBJECTIVE: To measure the anatomical position of the upper eyelid artery in the eyelid region, and to provide anatomical basis for adjacent flap grafting. METHODS: Twenty adult skull specimens were dissected, and a reference coordinate system was made based on the inner canthus connection for the X axis, and the center line for the Y axis. The red lactoprene was injected into the skull model via common carotid artery.The locations A-E of the upper eyelid artery in the eyelid area were measured. RESULTS AND CONCLUSION: The upper eyelid artery in the eyelid area was mainly from the supratrochlear artery and the supraorbital artery, generally paralleling to the X axis. The upper eyelid branch originated from the supratrochlear artery was located at the projection of the inner canthus, with a total length of 24.50 mm, and a diameter of 0.51 mm, extended to the outer canthus and the diameter of the vessel gradually reduced. The upper eyelid branch originated from the supraorbital artery was located at pupil and inner canthus junction 1/2 projection. The total length of the blood vessels was about 23-24.6 mm, and the diameter of the blood vessels was (0.55±0.05) mm. In the current study, we obtained the surface projecting of upper eyelid artery in the eyelid area by establishing the skull model of blood perfusion, which provides an anatomic basis for upper eyelid flap grafting.

ObjectiveWe report the clinical and pathological features of 8 Chinese myopathy patients with antibodies to the signal recognition particle(SRP).MethodsSerum myositis antibody profiles were tested with immunoblotting.Muscle biopsies were performed for histological,enzyme histochemical and immunohistochemical stainings.The first antibody in the immunohistochemical staining was mouse anti-human monoclonal antibodies including CD8,CD20,CD68,MHC- Ⅰ and CD31.ResultsEight cases showed stark positive of anti SRP antibody,3 of them with positive anti Ro-52 antibody.The muscle biopsies showed necrotic and regenerative muscle fibers associated with infiltration of macrophage,but scattered T lymphocytes in 2 patients.Two of them presented with fiber hypertrophy and proliferation of connective tissue.There were some fibers with positive MHC-Ⅰexpression.Capillaries were almost normal.Conclusion The muscle weakness of myopathy with antibodies to SRP presents as a chronic progressive course and could associate with lung involvement.Fiber necrosis and regeneration are the main myopathological features,which can mimic muscular dystrophy in some cases.

Objective To investigate the relationship between EphA2, EphrinAl and E-cadherin expressions and tumor stage and prognosis in pancreatic cancer. Method EphA2, EphrinAl and Ecadherin expressions were detected by immunohistochemistry in the tumor tissue and normal tissue specimens from 48 patients with primary pancreatic cancer. Results The expressions of EphA2 and EphrinAl were higher in the pancreatic carcinoma tissues than in the normal pancreatic tissues (P<0. 05). The E-cadherin expression was lower in the pancreatic cancer tissues than in the normal pancreatic tissues (P<0. 05). With decreasing histological differentiation, the expressions of EphA2 and EphrinAl in carcinoma tissues increased significantly (P<0. 05), while the E-cadherin expression decreased significantly (P<0. 05). The positive expressions of EphA2 and EphrinAl in the primary tumor significantly increased in stageⅢ and Ⅳ than in stage Ⅰ and Ⅱ (47. 9%vs 6. 25% , P<0. 05;47. 9% vs 8. 3%, P<0. 05), while the negative expression of E-cadherin was reversely correlated with these tumor stages (14. 6% vs 64. 6%, P<0. 05). Cox multivariate analysis showed that the clinical stage, EphA2 positive expression and E-cadherin negative expression were significantly associated with survival. Conclusion Abnormal expressions of EphA2, EphrinAl and E-cadherin were involved in the progression of pancreatic cancer and they were useful in predicting prognosis.