Objective To explore the effect of miniAd-ATP7B-GFP on the copper metabolism of skin fibroblasts of Wilson′s disease ( WD ) patients under high concentration copper medium.Methods Firstly, mini-adenovirus carrier containing human ATP7B gene was built and the mutations of 8 WD patients were detected.Fibroblasts from primary culture of skin of WD patients and normal human were cultivated 72 h in basic medium and medium with the copper concentration of C1(22.3μmol/L), C2(89.2μmol/L), C3 (156.1 μmol/L), C4 (245.3 μmol/L).Then the concentration of copper and protein was detected and copper/protein ratio was calculated.Secondly, miniAd-GFP ( miniAd-GFP group) and miniAd-ATP7B-GFP ( miniAd-ATP7B-GFP group ) were added into WD patients skin fibroblasts respectively, Wilson non-transfection group and normal group were set up as control, and C4 medium was used to culture the cells of four groups for 72 h and 96 h.Then the concentration of copper and protein was detected and copper/protein ratio was calculated.Results Five kinds of mutations were detected from 8 WD patients.The copper/protein ratio of WD patients and normal human in basic medium and the C1 -C3 groups had no statistically significant difference, but in C4 group (WD (1 871.6 ±209.2) ng/mg, normal group (1 267.2 ±188.3) ng/mg) the difference was statistically significant (t=6.075, P<0.01).C3((816.3 ±113.9) ng/mg) and C4 groups had statistically significant difference compared with the basic medium group ( ( 159.2 ± 38.6) ng/mg;WD:χ2 =31.493, normal group:χ2 =30.708, both P<0.01).The copper/protein ratio of 96 h group was higher than 72 h group.Compared with WD non-transfection (96 h:(2 731.2 ±188.7) ng/mg,72 h:(1 901.7 ±219.5) ng/mg) and normal groups, miniAd-ATP7B-GFP group had statistically significant difference both in 96 h ( ( 2 071.0 ±171.8 ) ng/mg ) and 72 h groups ( ( 1 495.5 ±161.4 ) ng/mg;72 h:F=20.130, 96 h: F=51.496,P<0.01).Conclusion MiniAd-ATP7B-GFP has partial improvement on copper metabolism of skin fibroblasts of WD patients under high concentration copper medium.

Objective To explore the effect of Tanshinone IIA on apoptosis and expression of Drp-1 and TRPM7 in a rat model of focal cerebral ischemia and reperfusion. Methods Rats were pretreated with high or low dose of tanshinone IIA before 2 h-focal cerebral ischemia plus 24 h-reperfusion. Cerebral blood flow in the middle cerebral artery was moni-tored during reperfusion. TTC, TUNEL and western blotting were used to detect the volume of cerebral infarction, apopto-sis and the protein expression of Drp-1 as well as TRPM7, respectively. Results Compared with control group, pretreat-ment with Tanshinone IIA could significantly down-regulate the expression of protein Drp-1 and TRPM7 (P<0.05), attenu-ate apoptosis (P<0.05), and reduce the volume of ischemia infarction. The volumes of right middle cerebral artery blood flow were(31.80%± 2.49%),(54.8%± 3.27%), and(58.8%± 3.03%)in controls, low-dose and high dose of tanshinone, respectively. Both low-dose and high-dose tanshinones improved cerebral blood flow. (tanshinone vs. control;all P<0.05). However, there was no statistical difference between low-dose and high-dose Tanshinone IIA groups in all measured out-comes (P>0.05). Conclusions Tanshinone IIA can inhibit ischemia-induced neuronal apoptosis and mitochondrial fission probably through improving cerebral artery blood flow and reducing the overexpression of Drp-1,TRPM7.

Objective To find the best time to remeasure temperature after cooling with evidence-based practice. Methods Used Johns Hopkins′ Evidence Based Practice Tool to find the best time to remeasure temperature after cooling. That tool included three steps which were establish problem, evidence synthesis and transfer evaluation. Results The best time to remeasure temperature after physical cooling was 30 minutes. The best time to remeasure temperature after drug cooling was 60 minutes. The best time to remeasure temperature after physical and drug combination cooling was 30 minutes after physical cooling and 60 minutes after using drug. Conclusion The best time to remeasure temperature after cooling was different because of the different cooling methods.

The research on clinical efficacy evaluation of traditional Chinese medicine (TCM) is a hot and difficult issue in the field of TCM. However, the overall level of current related research also needs to be improved. There are still a lot of academic controversies in the field of TCM clinical efficacy evaluation. The author believed that before conducting research, it is necessary to sort out core issues of TCM clinical efficacy evaluation, which in-clude the concept of TCM clinical efficacy and its evaluation. In recent years, Chinese scholars have conducted a series of studies on these issues and have some achievements. To think and answer these questions will play an important role for the ultimate resolution of problem of TCM clinical efficacy evaluation.

Through analyzing the current situation and coverage controversy of Chinese clinical trial insurance,this paper stated that the attending insurance rate in domestic clinical trials was entirely low.The sponsors,clinical trial institutions,investigators and insurance companies paid attention of different levels to clinical trial insurance.Therefore,the risk awareness of drug/medical device clinical trials should be enhanced.It is necessary to give impetus to clinical trial insurance system,during which all parties need to make a joint effort including government departments,ethics committees,sponsors,clinical trial institutions,investigators and insurance companies.

Objective · To explore the biological characteristics of platelet-rich fibrin (PRF) and its effects on proliferation and differentiation of adipose derived stem cells (ADSCs). Methods · The whole blood was collected from the forelimb vein of healthy beagles to prepare the PRF membrane, which were observed with optical microscope and scanning electron microscope. ADSCs were collected from the inguinaladipose tissue and were isolated and cultured. Identification of multi-directionaldifferentiation potential was performed. ADSCs were assigned to the PRF group and the control group, the former was treated with PRF in vitro. Cell proliferation wasmeasured with CCK-8. Osteogenesis induction was performed for two groups and the expression of genes associated with osteogenesis, including osteocalcin (OCN), osteopontin (OPN) and collagen I (Col- Ⅰ ), was measured with RT-PCR before induction and 4 and 7 days after induction. The alkaline phosphatase (ALP) activitywas measured 7 days after induction. Results · PRF is a milk white fibrin glue with elasticity and toughness. PRF can form loose and porous three dimensional network structure, which harbors lots of platelets and leucocytes. The cell proliferation activity was significantly higher in the PRF group than in the control group. After osteogenesis induction, the ALP activity and the mRNA levels of OCN, OPN, and Col- Ⅰ were significantly increased. Conclusion · PRF is a fibrin glue with three dimension network structure and contains lots of platelets, which can slowly release growth factors. PRF can promote the proliferation and osteogenic differentiation of ADSCs.

OBJECTIVETo prepare and observe the physicochemical properties of scaffold materials of heterogeneous deproteinized tissue-engineered bone.

METHODSDeproteinized bone was made through a series of physicochemical treatments in pig ribs and analyzed with histological observation, scanning electron microscopy, infrared spectrum, X-ray diffraction and energy dispersive analysis, Kjeldahl determination and mechanics analysis.

RESULTSInterstitial collagen fiber was positive and mucin was negative in deproteinized bone, but, both were positive in fresh bone. Deproteinized bone maintained natural pore network. Its pore size was 472.51 micromolar+/-7.02 micromolar and the porosity was 78.15%+/-6.45%. The results of infrared spectrum showed that collagen was present in deproteinized bone. Both fresh and deproteinized bone had curve of hydroxyapatite. The Ca/P ratios were 1.71+/-0.95 and 1.68+/-0.76 (P larger than 0.05), and the protein contents were 26.6%+/-2.23% and 19.1%+/-2.14% (P less than 0.05) in fresh and deproteinized bone, respectively. There was no significant difference of destruction load under compression and maximal destruction load between fresh and deproteinized bone (P larger than 0.05). The elastic modulus was higher in deproteinized bone than that in fresh bone (P less than 0.05).

CONCLUSIONSPhysicochemical properties and mechanic strength of deproteinized tissue-engineered bone meet the demands of ideal scaffold materials. But, its immunogenicity should be observed through further experiments for its clinical applications.

The objective of this study was to develop a method for the determination of taxol injection using near infrared transmission spectroscopy, turning out redetermination for injection from hospital pharmacy before using. Near infrared spectra (NIR) in the range of 12 000 approximation 4 000cm(-1) were recorded for the taxol injection manufactured during recent 24 months with different time. Calibration models were established using the partial least squares (PLS). Comparing different spectra pretreatments methods, dimension and spectra range, The study showed that spectra information cab be extracted thoroughly by delete a line method with dimension 6, spectra range 9002.5 approximately 4597.7cm(-1) ,standard error of the calibration sets(SEC) 0.035 and standard error of the prediction sets(SEP) 0. 059. Percent of prediction sets sample is less than +/- 2%. Results indicate that near infrared transmission spectroscopy method can be used to rapidly analyze the frequent and important drugs from hospital pharmacy.
Antineoplastic Agents, Phytogenic ; administration & dosage ; analysis ; Calibration ; Injections ; Paclitaxel ; administration & dosage ; analysis ; Spectroscopy, Near-Infrared ; methods

Objective:To observe the effects of self-designed Yiqi Huoxue Huayu Decoction on cardiac function and serum endothelin (ET-1) and matrix metalloproteinase-9 (MMP-9) levels in patients with chronic heart failure (GHF) complicated with atrial fibrillation.Methods:A total of 100 patients with GHF complicated with atrial fibrillation treated in our hospital from January 2019 to June 2021 were selected as the study subjects, and divided into experimental group and control group according to random number table method, with 50 patients in each group. Patients in both groups were given cardiotonic, diuretic, vasodilator and other conventional treatment, and patients in the experimental group added self-designed Yiqi Huoxue Huayu Decoction. TCM syndrome score, left ventricular end diastolic diameter (LVEDD), left ventricular end systolic diameter (LVESD) and left ventricular ejection fraction (LVEF), serum ET-1 and MMP-9 levels, clinical efficacy, safety and incidence of adverse reactions were observed and compared between 2 groups before and after treatment.Results:The total response rate was 82.0% (41/50) in the observation group and 62.0% (31/50) in the control group, and the difference was statistically significant ( χ 2=4.96, P=0.026). After treatment, the symptoms scores of palpitation, shortness of breath, fatigue, chest and hypochasm pain in the observation group were significantly significantly lower than those in the control group ( t=5.28, 5.29, 5.62, 5.42, P<0.01). After treatment, the LVEDD[(51.23±6.59)mm vs. (55.65±6.17)mm, t=3.46], LVESD[(43.10±4.76)mm vs. (45.99±5.31)mm, t=2.87], serum ET-1[(65.79±8.29)μg/L vs. (79.83±10.08)μg/L, t=7.61], MMP-9 [(175.86±24.81)ng/L vs. (189.49±26.13)ng/L, t=2.68] in experimental group were significantly lower than those in the control group ( P<0.05), while LVEF [(50.01±7.6)% vs. (46.25±6.96)%, t=2.57] was significantly higher than that of the control group ( P<0.05). There were no significant differences in GPT, GOT, UA, SCr levels between both groups before and after treatment ( P>0.05). There were no obvious adverse reactions betewwn both groups. Conclusion:The self-designed Yiqi Huoxue Huayu Decoction can relieve the clinical symptoms, improve the heart function and serum ET-1 and MMP-9 levels in patients with GHF complicated with atrial fibrillation safely.

Preeclampsia is the main cause of poor maternal-fetal outcomes. A series of cell and animal experiments, and a small number of clinical studies have shown that pravastatin can prevent and treat preeclampsia by regulating angiogenesis, increasing the expression of heme oxygenase, and stimulating the production of nitric oxide without any reported adverse effects during pregnancy. We review the latest progress on the mechanism, effect, and safety of pravastatin in the prevention and treatment of preeclampsia.