Objective To set up the quality control standard for Leaf of Chinese Holly; To provide basis for the utilization and development of Leaf of Chinese Holly.Methods Ten batches of Leaf of Chinese Holly samples from different habitats were collected, and the properties were described. The crosscutting and powder of leaf was under microscopic identification. Chlorogenic acid was set as reference substance to conduct thin-layer identification. Moisture, total ash, and acid-insoluble ash of the 10 batches were detected. HPLC was used to detect chlorogenic acid in Leaf of Chinese Holly.Results The properties and microscopic identification were described. Thin-layer chromatography method for chlorogenic acid in Leaf of Chinese Holly was formulated. Check items temporarily required that the moisture in Leaf of Chinese Holly should be less than 13%, total ash less than 13%, and acid-insoluble ash less than 5%. Content determination method for chlorogenic acid in Leaf of Chinese Holly was confirmed. It temporarily required that the content of chlorogenic acid should be more than 0.60%.Conclusion The established method can be used for the formulation of quality control of Leaf of Chinese Holly.

Objective To study the effect of glutamate on the expression of vascular endothelial growth factor (VEGF) mRNA and protein in cultured rat astrocytes. Methods Cultured rat astrocytes were randomly divided into 6 groups: control group (C), glutamate group (G), QA group (Q), DCG-IV group (D), L-AP4 group (L) and glutanmte-FMCPG gronp (G+M). Cells were cultured under nomoxic condition (95% air, 5% CO2). RT-PCR and ELISA methods were used to detect the expression of VEGF mRNA and protein in cultured astrocytes, respectively. G+ M group was preincubated with lmM MCPG for 30 min prior to the stimulation with glutamate. There were 7 time points at 0,4,8,12,16,24 and 48 h in each group except G+M group. Results The expression of VEGF mRNA and protein did not differ significantly among D group, L group and C group. Different from that in C group, the expression of VEGF mRNA and protein could be enhanced both in a dose-dependent and time-dependent manner in G group and Q group. Meanwhile, the enhanced expression of VEGF mRNA and protein in G group was completely suppressed by MCPG after 24 h. Conclusion Glutamate can increase the expression of VEGF mRNA and protein in cultured astrocytes, which may be due to the activation of group I metabotropic glutamate receptors in astrocytes.

Cinobufacino injection is purified from water extraction of the skin of Bufo bufo gargarizans, which has been widely used for various cancers in clinic with significant anti-tumor effects. Bufadienolides were regarded as the main active constituents of cinobufacino injection in previous reports. In present study, 6 bufadienolides were isolated and purified from Cinobufacino injection. Their structures were identified as 3-epi-ψ-bufarenogin (1), ψ-bufarenogin (2), 3-epi-arenobufagin (3), arenobufagin (4), 3-epi-gamabufotalin (5), and 3-oxo-arenobufagin (6), separately. Among them, 1 and 3 were new compounds, 5 and 6 were new natural products. Compounds 1, 2 and compounds 3, 4 were two pairs configuration isomers at C-3, separately.

Objective:To evaluate the efficiency of the tibial coronal alignment after total knee arthro-plasty with the extramedullary cutting system guided by the tibial tubercle and anterior tibial tendon,and to discuss the potential risk factors for the postoperative malalignment.Methods:A retrospective study of 212 primary unilateral TKA surgeries (in 188 patients)in our hospital between June to December in 2014 had been analysed.All the cases were performed by one surgical group.An extramedullary tibial cutting guide system had been used,with a landmark of one third inner-medial tibial tubercle as a proxi-mal anatomy reference,and anterior tibial tendon as a distal marker.The mechanical axis of lower ex-tremity in full-length X-ray was measured before surgery,and the tibial prosthetic coronal alignment was checked two weeks postoperation,evaluating the accuracy of this extramedullary cutting system guided by our method.Results:Since good alignment was defined as an angle within 3 degrees of the perpendicular to the mechanical axis,the tibial component positions were acceptable in 191 knees (90.1%),five knees were in valgus postoperative,and sixteen knees were in varus.There were seventeen(8.7%)in 179 knees with a preoperative varus presented malalignment after surgery,and four in 12 preoperative valgus kneess howed malalignment also,no statistical difference was found by Chi-square test (χ2 =2.778,P=0.096),which cannot define the relationship between the varus or valgus deformity preopera-tion and the malalignmentposition in tibial prosthesis after surgery.Twenty-two knees suffered a severe preoperative deformity as a varus or valgus angle larger than 20 degrees with absolute value of mechanical axis before surgery and tibial prosthetic coronal alignment were 21.95°(20.00°,26.90°)and 1.85° (0.10°,7.10°),showed a significant difference (Z =2.11,P =0.035 )compared with the data [10.65°(0.50°,19.80°)in preoperative mechanical axis and 1.10°(0.00°,4.60°)in the tibial pros-thetic coronal alignment]of 190 knees who presented a mild deformity before surgery(less than 20 de-grees),the result indicated the severe preoperative deformity might be a potential mal-alignment risk fac-tor within this cutting system in TKA surgery.Conclusion:The tibial coronal alignment after total knee arthroplasty could achieve good results with extramedullary cutting guide,by using one third inner-medial tibial tubercle and anterior tibial tendon as a proximal and distal anatomy marker.Postoperative varus might occur in this system,and tibial prosthetic malalignment appeared more often in the patients who suffered a severe deformity before surgery.

_ Objective_ To study whether quinoid dihydropteridine reductase ( QDPR ) expression level change can affect oxidative stress of NRK-52E renal tubular cells in a high glucose environment. Methods The NRK-52E model of overexpression, knockdown QDPR gene and respective control were constructed by lentivirus. All groups were given 5. 4 mmol/L and 30 mmol/L glucose culture medium respectively to imitate normal and high glucose condition. The level of superoxide anion ( O-2 ) was detected by flow cytometer dihydroethidium method. The protein expression level of superoxide dismutase 1 (SOD1)was tested by Western blot. Results QDPR over-expression can decrease O-2(P<0. 01)and SOD1(P<0. 05)levels in high glucose condition;QDPR knockdown increases O-2(P<0. 01) and does not change SOD1. Conclusion Under high glucose condition, overexpression of QDPR gene decreases NRK-52E cell oxidative stress. Knockdown QDPR gene increases NRK-52E cell oxidative stress. QDPR gene may influence the development of diabetic nephropathy by oxidative stress.

Antibody-drug conjugate (ADC) is a new class of therapeutics composed of a monoclonal antibody and small cytotoxin moieties conjugated through a chemical linker. ADC molecules bind to the target antigens expressed on the tumor cell surfaces guided by the monoclonal antibody component. The binding ADC molecules can be internalized and subsequently the toxin moieties can be released within the tumor cells via chemical and/or enzymatic reactions to kill the target cells. The conjugation combines the merits of both components, i.e., the high target specificity of the monoclonal antibody and the highly potent cell killing activity of the cytotoxin moieties. However, such complexities make the pharmacokinetic and metabolic studies of ADCs highly challenging. The major challenges should include characterization of absorption, distribution, metabolism and excretion, investigation of underlying mechanisms, assessment of pharmacokinetic- pharmacodynamic relationship, and analytical method development of ADC drugs. This review will discuss common pharmacokinetic issues and considerations, as well as tools and strategies that can be utilized to characterize the pharmacokinetic and metabolic properties of ADCs.

Objective: To explore effects of allogeneic hematopoietic stem cell transplantation (HSCT) in combination with infusion of endothelial progenitor cells (EPC) on bone marrow inflammatory injury. Methods: 6-8 weeks BALB/c (H-2K^d) mice after lethal dose of irradiation (TBI) were subjected to allogeneic bone marrow transplantation (BMT group) or co-transplantation of EPC (EPC group) . Samples of bone marrow cells of mice in each group on days 7,14,21,28 after transplantation were obtained to detect EPC cultural and cell chimeric rates by flow cytometer. Mice were sacrificed on days 7, 14, 21 and 28 post HSCT to analyze bone marrow pathology by H&E staining, the infiltration of macrophages and neutrophils by Western blot, validation expression levels of inflammatory complexes nlrp1、nlrp6 and its downstream molecules casepase-1 by Q-PCR and Western blot. Results: Cell chimeric rate on day 7 after transplantation in EPC group[ (91.65±2.77) %] was significantly higher than in BMT group[ (83.69±1.26) %]. Alleviated osteomyelitis injury and inflammatory cell infiltration in EPC group were observed when compared with BMT mice. Also significant reductions of the levels of nlrp1、nlrp6、casepase-1 transcription complexes in EPC mice were noted when compared with BMT ones. Conclusion Co-transplantation of HSC and EPC could alleviate inflammatory cell infiltration and activation of the complex to promote the repair of bone marrow.

Objective To investigate the protective role of Sestrin2 overexpression in hypoxia and re-oxygenation injury (H/R) injury of hippocampal neurons and its mechanism in rats.Methods Neurons were enzymatically isolated from hippocampi of newborn Sprague-Dawley rats (less than 24 h old) and culturedinvitro. These neurons were randomly assigned into 4 groups (n=20) using a random number table: control group, H/R group, vector group and Sestrin2 overexpression group. The hippocampal neurons were seeded in 6-well plates at a density of 2×104 cells/mL; neurons in the latter two groups were transfected with lentiviruses containing empty vector andSestrin2overexpressed genes, respectively; the hippocampal neurons in the later three groups were subjected to oxygen-glucose deprivation (OGD) for 6 h followed by restoration of O2 supply for 20 h. The reactive oxygen species (ROS) content was detected by Reactive Species Assay Kit, and the ATP concentration was detected by ATP Assay Kit. Cell apoptosis rates were measured by flow cytometry. The levels of Sestrin2, dynamin-related protein 1 (Drp1), Fis1, and apoptosis-related proteins Bcl-2, Bax and cytochrome C (Cyt C) were measured by Western blotting. The ratio of Bcl-2 to Bax was calculated. The ultrastructure of mitochondria was observed by transmission electron microscopy.Results As compared with control group, H/R group had significantly lower ATP concentration, Bcl-2 protein expression and ratio of Bcl-2 to Bax ([11.15±0.42] nmol/mg proteinvs. [5.30±0.39] nmol/mg protein; 2.20±0.26vs. 0.91±0.02; 6.46± 0.41vs. 1.04±0.05), statistically higher average fluorescence intensity of ROS and cell apoptosis rate (152.41±17.38vs. 1530.00±14.69; 3.77％±0.74％vs. 56.57％±1.35％), and significantly higher protein levels of Sestrin2, Drp1, Fis1, Bax and Cyt C (0.66±0.06vs. 1.11±0.20; 0.48±0.03vs. 1.16±0.07; 1.14± 0.09vs. 2.47±0.09; 0.34±0.03vs. 0.88±0.04; 0.17±0.03vs. 0.30±0.03,P<0.05); what's more, the structure of mitochondria was obviously destroyed in I/R group. As compared with H/R group, Sestrin2 overexpression group had significantly increased ATP concentration, Sestrin2 and Bcl-2 protein expressions and ratio of Bcl-2 to Bax ([8.95±0.27] nmol/mg protein; 2.67±0.07; 1.80±0.19; 3.95±0.28), significantly lower average fluorescence intensity of ROS and cell apoptosis rate (337.27±15.32; 10.33％±2.60％), and statistically lower protein levels of Drp1, Fis1, Bax and Cyt C (0.43±0.02; 1.11±0.08; 0.45± 0.02; 0.17±0.02,P<0.05); the structure of mitochondria was relatively completed in Sestrin2 overexpression group.Conclusion Sestrin2 overexpression can inhibit mitochondrial fission, reduce accumulation of reactive oxygen species, improve mitochondrial energy metabolism and block mitochondria mediated apoptosis pathway, thereby alleviating I/R injury of rat hippocampal neurons.

Objective: To investigate the usage of medical and health care APP among female college students, and to understand possible influencing factors, so as to provide reference for medical health care behaviors and personal health among college students. Methods: A stratified random sampling method was used to select 891 female college students from 3 colleges in Weifang. A questionnaire survey on their concentration on their own health, the knowledge, cognition and behavior of medical and health care APP, and Logistic regression analysis was used to analyze the influencing factors. Results: There were 55.4% female college students who had used medical and health care APP, and most of them (290) had used fitness and weight loss APP. There was statistical significance of the different usages of students of different degrees, majors and grades(χ2=26.55, 7.72, 13.96, P<0.05). Logistic regression analysis showed that educational background, annual medical costs, awareness of medical and health care APP and attitude towards APP prospect were the factors influencing the utilization of medical and health care APP (OR=0.44，0.69，1.36，1.23，P<0.05). Conclusion The usage rate of medical and health care APP is general, and sustained utilization rate is low, the APP should enrich menstrual period consumption and figure report, add video recommendation, interactive dating, and other functions, improve female students medical and health care APP cognition, female college students to change health care APP attitude.

Objective To investigate the influence of curcumin and its analogue H8 on glucose and lipid metabolism disorder in db/db mice. Methods The type 2 diabetes mouse model (db/db mice) was intragastrically administrated with curcumin and analogue H8 for 8 weeks.The blood biochemical indexes were measured.The expression of PEPCK and G6Pase mRNA was detected by real-time PCR in liver tissues.The expression of PEPCK and G6Pase protein was detected by Western blotting. Results Curcumin analogue H8 reduced blood glucose and lipids in db/db mice (P<0.01) and improved liver function related enzymes significantly.The levels of PEPCK and G6Pase mRNA in db/db mice were significantly decreased (P<0.01) and the expression levels of PEPCK and G6Pase protein were significantly decreased(P<0.01). Conclusion Curcumin analogue H8 improves the glucose and lipid metabolism disorder in db/db mice,and it is related to inhibiting the expression of PEPCK and G6Pase gene and protein.