Objective: To establish the assay method of 2,3,5,4′-tetrahydroxystilbene-2-O-?-D-glucoside in Jiangzhitongmai Tablet (Radix Polygoni Multiflori, Rhizoma Alismatis, etc.). Methods:HPLC conditions were as fellows: Chromasil C 18 column, a mobile phase of CH 3CN-H 2O(25∶75), and the wavelength at 320nm. Results:2,3,5,4′-tetrahydroxystilbene-2-O-?-D-glucoside was linear within the range of 0.044～ 0.7 ?g with a correlation coefficient of 0.9999. The average recovery was 98.54%. Conclusion:The method is simple, swift, accurate and practical in quality control of Jiangzhitongmai Tablet.

Objective To investigate the expression patterns of fibroblast growth factor 23 (FGF23) in osteoblast and its responses to calcium, phosphate, exogenous PTH and 1,25(OH)_2D~3. Methods The primary rat calvarial osteoblasts were cultured in MEM medium which containing 10% FBS, then were harvested when cells were in half-confluence, confluence, osteoid deposition and osteoid mineralization stages respectively. The procedure was monitored under microscopy. Total RNA was extracted from cells according to the Trizol procedure. FGF23 mRNA levels were determined by Real-time PCR. Further, the confluent osteoblasts were treated with 3.2 mmol/L CaCl_2, 4.4 mmol/L β-glycerophosphate, 10~(-9) mol/L rhPTH(1-34) and 10~(-8) mol/L 1,25(OH)_2D_3 respectively for 3 days, and same volume of the medium was added as the control. The gene expressions were determined by Real-time PCR. Results FGF23 expression was transiently up-regulated at cell confluent stage and down-regulated after that. The FGF23 mRNA levels were 7.5-fold higher in confluent cells compared with that in half-confluent cells (P<0.001). The markedlly stimulating effect (about 16 times) on FGF23 expression was stimulated by exogenous 1,25(OH)_2D_3 treatment while no significant effect was found on FGF23 mRNA levels by CaCl_2,β-glycerophosphate, and rhPTH(1-34) treatments when compared with the control. Conclusions The FGF23 expression in osteoblast is developmental stage-related and its powerful stimulator is 1,25(OH)_2D.

Objective To observe the earlier pathological character and mechanism of radiation osteonecrosis in femoral head, in order to provide evidences for the earlier diagnosis and prevention of radiation osteonecrosis of femoral head. Methods Single femoral head of rats were irradiated singly with 30 Gy of 137 Cs γ-ray. The rats were executed after 2, 6 and 12 weeks, then the femurs were stained with HE and histopatholngical changes were observed by light microscope. The bone marrow mesenchymal stem cells (BMSCs) were cultured after 2 weeks and its proliferation and the colony formation were observed. The rats were endo-perfused with microfili contrast medium 12 weeks later, and the 3-dimensional structure of capillaries by Micro-CT was re-estabhshed to detect the pathological changes of capillaries after irradiation. Result The irradiated femur showed deranged cbondrocyte, decreased osteocyte, shrinking nucleus, increased empty bone lacuna and reduced bone trabocnla (P < 0.05). Micro-CT showed the discontinued small vessels and absence(6.65 %) capillaries in irradiated femur were obviously less than those of the unirradiated (12.3 %)(P < 0.001). The proliferation of BMSCs was slowed, the number of colony in irradiated group (10 %) was less than that of control (21 %) (P < 0.001). Conclusions The preliminary histopathological changes of osteoradionecrosis on femoral head could be increased the empty bone lacuna, and the bone lacuna above 30 % was the sign of the earlier period of osteoradionecrosis. The osteonecrosis of femoral head induced by radiation is not only correlated to the damages to the bone, but also to the damages to BMSCs and capillaries.

To study whether the late-acting co-stimulatory molecules ICOS can suppress the apoptosis and sustain the survival and proliferation of T cells through the survivin pathway, ICOS signals deficient T-cells were infected with adenovirus carried survivin gene, other T-cells were given ICOS co-stimulatory signals, then infected with adenovirus carried dominant-negative mutant survivin gene. Apoptosis and proliferation were determined by TUNEL and CCK-8 respectively. The results show that engagement of ICOS signal increased the expression level of survivin significantly. Survivin can sustain co-stimulatory deficient T cells survival and suppress the apoptosis. Mutant survivin inhibits ICOS signal positive T cells survival and increase its apoptosis. Late-acting co-stimulatory molecules ICOS can suppress the apoptosis and sustain the survival of T cells through the survivin pathway.

BACKGROUND:The homing ability of mesenchymal stem cells is closely associated with the effects of celltransplantation. Clarifying the mechanism of chemotaxis and migration wil contribute to enhance the clinical application of mesenchymal stem cells. OBJECTIVE:To investigate the effect of Cdc42 in the homing of human umbilical cord mesenchymal stem cells. METHODS:First, mesenchymal stem cells were isolated from human umbilical cord, and co-cultured with tumor necrosis factorα, interleukin-1β, and transforming growth factorβ. Western blot assay was used to test the level of Cdc42. Besides, Cdc42 siRNA was synthesized by chemical method to transfect the cells, and cellmigration and adhesion were measured by Transwel and Matrigel separately. Meanwhile, the activity of signal molecule, extracellular regulated protein kinase 1/2, was evaluated by western blot. RESULTS AND CONCLUSION:The results indicated that the inflammation factors induced the highly expression of Cdc42 in human umbilical cord mesenchymal stem cells, almost double level to controls. siRNA notably inhibited the migration and adhesion of human umbilical cord mesenchymal stem cells through Cdc42 down-regulation, and the extracellular regulated protein kinase 1/2 and phosphorylation form were also decreased simultaneously. In a word, we speculate Cdc42 plays a role in the chemotaxis of human umbilical cord mesenchymal stem cells in vitro.

OBJECTIVE:To provide reference for the rational use of Potassium chloride injection and the management of high-risk drugs. METHODS:A total of 1 065 prescriptions containing Potassium chloride injection during the first half year of 2014 were analyzed retrospectively according to“Rules for Comment on Prescriptions”. RESULTS:The qualification rate of pre-scription was 95%. The irrational prescriptions accounted for 5%. The main problems included unreasonable route of administra-tion,unreasonable selection of solvent,incompatibility with TCM injection and other types of injections as well as the risk of Potas-sium chloride injection combined with a few oral drugs. CONCLUSIONS:The defect still exist in the management of high-risk drug aspotassiam chloride injection in our hospital,so that the hospital should set up high-risk drug prescription special review sys-tem and emergency plan which is the effective way for avoiding the drug risk of high-risk drugs.

In the presence of silver ion, Mn2+ could be electro-oxidized to potassium hypermanganate in phosphoric acid solution, which could effectively react with pyrocatechol in acid solution and luminol in sodium hydroxide solution to produce chemiluminescence. On the basis of this, a novel indirect approach for the detection of Mn2+ was established. The effect of silver ions on the electrochemical oxidation of Mn2+was studied. when 1. 5 ×10-5 mol/L Ag+ and 0. 01 mol/L phosphoric acid solution were used in the process of electrochemical oxidation, the CL intensity could be up to the maximum value after the above solution was electrolyzed for 2 min. The relation of CL intensity and Mn2+concentration in the solutions at different pH and the selectivity were also investigated. when the pyrocatechol was used as luminescent reagent in the acidic medium, the CL intensity was linearly to the Mn2+concentration in the range of 1. 82×10-7-7. 27×10-5 mol/L with excellent selectivity. Common ions had little interferences in the determination of Mn2+. The method was successfully applied to the determination of Mn2+ in surface water and drinking water with satisfactory results.

Objective To explore the effects of daidzein (DA) on the expressions of estrogen receptors (ER) and peroxisome proliferator-activated recepor γ (PPARγ) in osteoblasts and the influence of estrogen on these effects.Methods A mouse osteoblastic cell line MC3T3-E1 cultured in α-MEM containing 2％ FBS was treated by 0.1 and 10 μmol/L DA.ER antagonist ICI182780 and PPARγ antagonist GW9662 in 0.1 μmol/L was added as required,and an equivalent amount of phosphate buffer solution (PBS) was used as control.For the study on estrogen effect,the cells were treated by DA in the serum-free medium with or without 10 nmol/L 17β-estradiol (E2).The expressions of ERa,ERβ and PPARγ were determined by real-time RT-PCR and Western blot analysis,respectively.Results DA inhibited ER,expression but stimulated PPARγ expression in the cells at the concentration of 0.1 and 10 μmol/L.The down-regulation of ERα by DA could be blocked by ICI182780,whereas the up-regulation of PPARγ could be repressed by GW9662 in transcription levels.Furthermore,the inhibitory effect of DA on ERβ expression was markedly enhanced,while its stimulatory effect on PPARγ expression was almost lost in serum-free medium with 10 nmol/L 17βestradiol as determined by real-time RT-PCR.Conclusions Besides its direct roles in ERs and PPARγ mediated gene transcriptions,DA could exert indirect effect on cellular pharmacological responses by altering ER and PPARγ expressions.The predominant influence on receptors expression probably involved in the time-related biphasic effects of DA on osteogenesis,which was supposedly influenced by estrogen level.

To explore the effect of PluronicF-127 and Vascular endthelial growth factor(VEGF) delivery system on the survival of the grafted fat, we divided fat harvest under the same condition into 4 groups. One group served as blank control; the other 3 groups served for experiments with respective to DMEM containing 20 ng/ml VEGF; DMEM containing 30% Pluronic F-127; DMEM containing 20 ng/ml VEGF and 30% Pluronic F-127, and then we transplanted the 4 groups of fat, subcutaneously, on the back of 3 groups of BALB/c nude mice (8 mice per group; injecting 4 points per mouse; 0.2 ml per point). At 3 weeks, 6 weeks and 12 weeks, we dissected the fat grafts, measured their weight retention, and put them in histopathologic examination with the use of HE and CD34 staining. And we compared the weight retention and microvessel density (MVD) of each experiment group versus those of control group. The relation between adipose cell and PluronicF-127 was observed through electron microscope. The results reveal that the MVD and weight of pluronicF-127 and VEGF of the experiment groups are significantly greater than those of other groups. The PluronicF-127 and VEGF composite delivery system can significantly improve the blood circulation for fat transplantation, and increase the survival rate of grafted fat.
Adipose Tissue ; cytology ; transplantation ; Animals ; Drug Delivery Systems ; Female ; Graft Survival ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Poloxamer ; administration & dosage ; Random Allocation ; Vascular Endothelial Growth Factor A ; administration & dosage

Objective To investigate the isolation rate, distribution and trend of nontuberculous mycobacteria (NTM) in Wenzhou during 2014 to 2016.Methods Sputum or alveolar lavage specimens of patients with suspected pulmonary tuberculosis were collected for mycobacteria culture from January 2014 to December 2016.Mycobacterium culture positive strains were further identified by gene chip, 16S rRNA and hsp65 gene sequencing.SPSS 19.0 software was used to analyze the data.Results After excluding repetitive strains, 3 295 mycobacteria strains (MTB) were isolated from respiratory specimens, included 3 032 mycobacterium tuberculosis complex strains, 238 NTM strains, 20 Gordon genera strains, 3 Nocardia genera strains and 2 Tsukamurella genera strains.The proportion of NTM among confirmed mycobacteria was 8.5％ (86/1 006), 6.7％ (72/1 079) and 6.8％ (80/1 185) in 2014, 2015 and 2016, respectively (x2 =2.459,P ＞ 0.05).The overall prevalence of NTM was 7.3 ％ (238/3 270).There were 15 species of NTM, and the most common NTM strain was Mycobacterium intracellulare (52.5％,125/238), followed by Mycobacterium abscessus (22.7％, 54/238) and Mycobacterium avium (10.1％, 24/238), other species were only accounted for 14.7％ (35/238).The ranking of Mycobacterium avium went up rapidly from the fifth in 2014 to the second in 2016 (x2 =18.259, P ＜ 0.01), while proportion of Mycobacterium abscess, dropped from 34.9％ (30/86) in 2014 to 17.5％ (14/80) in 2016 (x2 =7.335, P＜0.01).Among patients from whom the NTM strains were isolated, 56.7％ (135/238) were male and most of them were aged 45 years or above (79.8％, 190/238).Conclusions In the past three years, the trend of NTM isolation rate in Wenzhou is steady.The most prevalent NTM species is Mycobacterium intracellulare, followed by Mycobacterium abscessus and Mycobacterium avium.Mycobacterium avium shows a continuously upward trend, while the separation of Mycobacterium abscessus shows a downward trend.