OBJECTIVE:To determine the content of paeonol in Cynanchum paniculatum by three-dimensional fluorescence coupled with ATLD algorithm. METHODS:The trilinear model was established. The principle of least square was used for ATLD. The component value was fitted by core consistency diagnosis,and inner filter effect was corrected by mathematical correction method. Fluorescence scanning condition included excitation wavelength of 250-400 nm,emission wavelength of 410-600 nm, wavelength interval of 5 nm,slit width of 5.0/5.0 nm,scanning speed of 1200 nm/min. Determination and absorption spectrum condition included scanning wavelength of 250-600 nm,scanning speed of 600 nm/min;Al(Ⅲ)was used as the sensitizer to in-crease the fluorescence intensity of paeonol. The content of paeonol was determined by HPLC. RESULTS:The linear range of pae-onol was 0.132-1.188 μg/mL(r=0.9999). RSDs of precision,stability and reproducibility tests were less than 3.0%. The recover-ies were 100.1%-104.7%(RSD=2.39%,n=6). The analysis spectrum of paeonol was almost completely overlapped with the actu-al spectrum. The results of HPLC method are very similar to those of ATLD algorithm. CONCLUSIONS:The three-dimensional fluorescence coupled with ATLD algorithm is simple,rapid,efficient and accurate,and can be used for the qualitative and quantita-tive analysis of complex system.

Objective:To investigate the effect of short term and intensive insulin pump on the blood lipid and glucose levels of patients with type 2 diabetes.Methods:76 patients with type 2 diabetes were selected and divided into two groups according to different administration.The control group (38 cases) was given routine insulin aspart treatment.The observation group (38 cases) was treated by insulin pump.The blood and lipid indexes and ADL score before treatment,at 1 week and 2 weeks after treatment were compared between two groups.Results:After treatment,the levels of FPG,HbAlc,TC,TG and LDL-C were lower than those before treatment and the level of HDL-C was increased in both groups.The levels ofFPG,HbAlc,TC,TG,LDL-C of observation group were lower than those of the control group,and the level of HDL-C of observation group was higher than that of control group (P＜0.05).At 1 week,2 weeks after treatment,the ADL scores of both groups were significantly improved,and the ADL score of observation group was higher than that of the control group (P＜0.05).Conclusion:Short term intensive insulin pump could significantly improve the blood glucose and lipid metabolism disorders in patients with type 2 diabetes,and improve the ability of daily life.

Objective: Orbital malignant tumor tends to infiltrate the skin,muscle and maxillae antrum.Its treatment requires enlarged surgery,which may cause bone exposure and make it difficult to repair the big wound.The purpose of this study was to explore the method of using the super-width forehead musculocutaneous flap to repair the postoperative wound.Methods: Based on the features of arterial anastomisis,we used super-width forehead musculocutaneous flaps,with the supraorbital and supratrochlear arteries as the pedicle,to repair the wound following surgical excision of orbital malignant tumor in 6 patients.Results: The wounds of the 6 patients were 5?5 to 8?9 cm2 in size,with the related part of the skull exposed.The musculocutaneous flaps survived in all the patients and all the wounds were successfully repaired in the primary stage.Conclusion: It is a good option to repair the defect left by orbital malignant tumor excision using super-width forehead musculocutaneous flaps with the supraorbital and supratrochlear arteries as the pedicle.

Objective To reconstruct the perfusion parameters of murine hindlimbs peripheral tissue by using fluorescence reflectance imaging technique. Methods BALB/c mice were injected with intravenous bolus injection of indocyanine green (ICG) (10μg) into the tail vein. Time-series fluorescence intensity images were obtained for 200 s immediately after the injection. After the serial imaging, silhouette images of the mice were taken under white light to obtain the region of interest (ROI) of the murine hindlimbs. Bi-exponential model was applied to analyze the dynamic fluorescence parameters and the peripheral tissue perfusion parameter images were reconstructed. Results The fitted perfusion curves obtained from bi-exponential model were in good agreement with the measured ones. The parametric images which reflected the vascular sufficiency of murine hindlimbs were reconstructed. Conclusions A novel method for parametric images of murine hindlimbs peripheral tissue blood perfusion is proposed in this paper, which is noninvasive with higher resolution and little damage to biological tissues.

Objective To determine the expression of Notch pathway receptors (Notch1 and Notch4) and ligands (Jagged1 and Dll4) in cutaneous malignant melanoma (CMM) tissues,and to preliminarily explore the role of the Notch signaling pathway in the pathogenesis of CMM.Methods Immunohistochemical study was performed to determine the expression pattern and intensity of Notch1,Notch4,Jagged1 and Dll4 in 40 paraffin-embedded CMM specimens and 15 paraffin-embedded pigmented nevus specimens.Statistical analysis was carried out by chi-square test and Spearman rank correlation analysis with the SPSS 21.0 software.Results Notchl was detected in 31 (77.5％) of 40 CMM specimens,as well as in 3 of 15 pigmented nevus specimens,and the positive rates significantly differed between the two groups (x2 =15.281,P ＜ 0.001).However,no significant difference in the expression intensity of Notch1 was observed between 18 in situ melanoma tissues and 22 invasive melanoma tissues (x2 =0.631,P =0.427).In addition,the positive rates of Notch4,Jagged1 and Dll4 were also significantly higher in the CMM group than those in the pigmented nevus group (all P ＜ 0.05),and the expression intensity of Notch4,Jagged1 and Dll4 significantly differed between in situ and invasive melanoma tissues (all P ＜ 0.05).In CMM tissues,the expression of Notch1 was positively correlated with that of Jagged1 (rs =0.350,P =0.027) and Dll4 (rs =0.562,P ＜ 0.001),while the expression of Jaggedl was negatively correlated with that of Dl14 (rs =-0.734,P ＜ 0.001).Conclusion Abnormality of the Notch signaling pathway may be involved in the pathogenesis of melanoma,but further researches are still needed to elucidate the detailed mechanism.

OBJECTIVE： The compound nucleic acid granules were prepared， observed for their therapeutic effects， and determined for their pharmacological actions METHODS： Quality control was used in their preparation The therapeutic effects were observed on 22 patients with coronary heart disease of stable angina pectoris with hyperlipemia who were chosen according to clinical diagnosis standard RESULTS： （ 1） Improvment of clinical symptoms for angina pectoris： satisfactory 22 7％ ， improved 68 2％ ， failure 9 0％ ； （ 2） Blood lipid analysis： cholesterols and triglycerides were improved significantly after administration of the preparation（ P0.05;P<0.01).CONCLUSION:The compound nucleic acid granules have marked effects on coronary heart disease, angina pectoris and hyperlipemia.

Objective By analyzing the epidemiological characteristics and trends of brucellosis in Ji'nan City,to provide a scientific basis for further prevention and control of the disease.Methods Serum samples of suspected brucellosis patients were collected from every county (city,district) of Ji'nan City from 2008 to 2014,and these samples were screened by Rose-bengal plate agglutination test (RBPT).Then positive samples were confirmed by Standard tube agglutination test (SAT),and experimental data and results were analyzed by statistics.Results Totally 2713 serum samples of suspected brucellosis patients were tested,and 200 were positive by SAT.Temporal distribution:the serum antibody positive specimens were detected each month throughout the year,and the results showed clear seasonal prevalence by concentrating from March to September (82.07％,151/184).Population distribution:the serum antibody positive rate of males was higher than that of females,the ratio was 2.08:1.00,and the cases were found among 15 to 75 years old,while concentrated from 30 to 65 (67.00％,134/200).Spatial distribution:except Huaiyin District,positive samples were detected in all other 10 counties (cities,districts) of Ji'nan City,most of them (92.5％,185/200) in Shanghe,Zhangqiu,Changqing,Shizhong and Licheng counties (cities,districts).Conclusions Brucellosis epidemic in Ji'nan is serious,and cases have occurred mostly in spring,summer and autumn.It is recommended that health propaganda and education should be strengthened in order to enhance selfprotection awareness of the exposed population,and then improve the physical examination for early discovery,early reporting and early treatment.At the same time,it is important to cooperate with the Department of Livestock to prevent and control brucellosis.

Objective To study the Dai medicine Yapa Tang inhibition on mice Lewis lung carcinoma growth and metastasis,and to explore the impact of the drug on mice immune function. Methods Subcutaneous injection LLC(Lewis lung carcinoma cells)in C57BL/6 mice’s right armpit was to establish rat models. 60 successful tumor-bearing mice were selected and thenafter inoculating LLC were randomly divided into 6 groups, namely the control group,Yapa Tang low-dose group,Yapa Tang mid-dose group, Yapa Tang high-dose group,cyclophosphamide group and Yapa Tang mid-dose combining cyclophosphamide group.Then,after inoculated 3 days,each group was administrated with medicines.The first four groups wereintragastricly administrated with 0.9% saline, low-dose, mid-dose and high-dose Yapa Tang with0.4ml respectively;the cyclophosphamide group was administrated with cyclophosphamide,and Yapa Tang mid-dose combining cyclophosphamide group was administrated with middle dose Yapa Tang and cyclophosphamide. After 21days, blood was taken from eyeballs. Tumor tissue, spleen, thymus were gotten and weighted to calculate inhibitory rate,spleen and thymus index, enzyme-linked immunosorbent assay (Elisa) was adopted to detect Interleukin-2(IL-2), Interleukin-10(IL-10)and their contents. Results The tumor weight of Yapa Tang high-dose group, cyclophosphamide group and Yapa Tang mid-dose combining cyclophosphamide group was 3.46±0.39, 2.39±1.04 and 2.30±0.76 respectively,all lower than the control group, which was 5.21±0.50, showing significant difference(P<0.05), with the inhibition rate were33.69%, 54.12%, 56.00%. The thymus and spleen index of Yapa Tang low-dose group, middle-dose group and high-dose group was2.16±0.69, 2.24± 0.76, 2.23 ± 0.63, 16.82 ± 3.14, 15.82 ± 1.72, 17.08 ± 3.65, significantly higher than that of the control group,which was1.94±0.6, 15.17±3.53. The IL-2, IL-10 level of control group were(883.54±181.49)ng/L, (1 106.86±343.79)ng/L. Yapa Tanggroupsshowed an increase in IL-2(1 732.29±100.52)ng/L, (1 813.33± 168.32)ng/L, (2 275.63 ± 394.76)ng/L and the decrease in level of IL-10, respectively were(834.02 ± 271.97)ng/L, (636.83±270.56)ng/L, (682.08±147.85)ng/L, with significant difference. Conclusion Yapa Tang can inhibit lewis lung cancer grow and reduce the number of lung metastases, regulate immune cytokines IL-2 and IL-10, and promote the immunity of tumor-bearing mice.

Objective To investigate the effects of transplantation of allogenic adipose-derived stem cells (ADSCs) on bone metabolic markers and bone histomorphometry in ovariectomized (ovx) female rats.Methods Thirty female nonpregnant Wistar rats were randomly divided into two groups:the ovx group (n=20) and sham operated group (n=10).Three months after operation,two rats in each group were randomly sacrificed for ovariectomized osteoporosis model assessment.The serum levels of bone-specific alkaline phosphatase (BALP) and tartrate-resistant acid phosphatase (TRACP) were detected by enzyme-linked immunosorbent assay (ELISA).The pathological changes of femur were observed by HE staining.After animal model was developed,the rest of the rats in the ovx group were further subdivided into two groups:a control group and a treatment group.Each rat in the treatment group was treated with ADSCs via tail vein injection once a week,while each rat in the control group was injected with the same dose of saline.After eight weeks of treatment,the serum levels of BALP and TRACP were measured by ELISA.The expression of osteoprotegerin (OPG) and receptor activator of nuclear factor κB ligand (RANKL) in femur was detected by immunohistochemical staining technique.In addition,the pathological changes of bone were observed by HE staining.All data were analyzed by one-way ANOVA and LSD-t test using SPSS 13.0 software.Results Compared with the sham operation group,the rats in the ovx group showed lower serum level of BALP and higher content of TRACP.Morphology analysis showed that the trabeculae were more narrow,broken more and less in number in the ovx group.In addition,the treatment group showed higher serum level of BALP [(49±7) mU/ml,(42±5) mU/ml,respectively] and lower TRACP [(31±5) μg/L,(37±5) μg/L,respectively] content than the control group (P＜0.05).Immunohistochemical staining demonstrated that the treatment group had higher OPG expression (0.011 3±0.001 6) in femur than the control group (0.007 7±0.001 5),however,no significant difference in RANKL protein expression level was detected.Moreover,treatment with ADSCs induced extensive new trabecular bone formation and reduced potential tissue damage in the trabecular bone when compared with the control group.Conclusion Transplantation of ADSCs via tail vein injection protects against ovariectomy induced bone changes in rat model.The results of this study suggests that this treatment may potentially be used in the treatment of osteoporosis.

Objective To identify the animal drug of Cervus elaphs and C. nippon from origin of deers. Methods To extract DNA from deer blood and hairy antler of 11 species of deers such as C. elaphus, C. nippon and so on, and to gain the mitochondrial 12S rRNA gene fragment using the general primers of L1091 and H1478. Based on the sequence multialinement of 11 species deers above gene fragments, designing the couples of special difference primers and identifying C. elaphs and C. nippon. Results 12S rRNA Gene fragments can distinguish different deers well. The couple of primers (EP-1/H1478 and EP-2/H1478) PCR can effectively identify C. elaphus and C. nippon. Conclusion Special primer PCR is suitable for the identification of valuable Chinese medicinal materials, such as C. elaphus and C. nippon.