Objective:To study the impact of circadian variations on the efficacy of urokinase (UK) therapy in patients with acute myo- cardial infarction (AMI). Methods:Ninety-five AMI patients presenting with chest pain≤3 hours were given UK theerapy.They were divided into two groups according to their symptom onset time:late morning group (6:01—12:00,n=42) and other time group (12:01—6:00 next day,n=53).We obtained elbow venous blood samples at 3 time points:before UK therapy,30 and 60 minutes after UK ini- tiation respectively.Serum levels of AT-Ⅲ,PAI-1 Ag,PAI-1 activity,D-D,?_2-APL Ag were examined.The repatency of culprit vessels was also determined 120 minutes after UK administration. Results:①UK therapy in the late morning group(23 out of 42 patients) had lower repatency rate compared with the other time group(40 out of 53 patients) (P＜0.05).②The late morning group had a higher serum levels of PAI-1 Ag before UK thera- py (39.822?4.737 versus 37.406?4.696,P＜0.05).③The serum level of PAI-1 antigen at aforementioned three time points was 36.054?4.456,35.242?4.143 and 35.355?4.135 in reperfused patients compared with 42.7822?3.277,42.398? 4.565 and 42.395?5.223 in nonnperfused patients.The activity of PAI-1 was 15.234?2.671,13.256?4.532 and 11.830? 3.905 in reperfused patients versus 17.0743?3.665,16.447?4.469 and 16.928?4.248 in nonreperfused patients at three time points.Both of PAI-1 antigen and its activity was consistently lower in reperfused AMI patients as compared with the nonper- fused patients before and 30,120 minutes after UK therapy(P＜0.05).There was no significant difference in the serum levels of ?_2-APL Ag,D-D,and AT-Ⅲ activity between the two populations (P＞0.05). Conclusion:Our data showed circadian variations have an impact on the efficacy of UK therapy in patients with AMI,which suggests that there may exist a phenomenon of “morning resistance to thrombolysis therapy” due to increased level of PAI-1 Ag and activity.

Objective To evaluate the effect of the patient's preoperative visit to the operating room on the patient's apprehension before operation.Methods One hundred and Sixty patients aged 16-64 yr with fear visual analog scale (FVAS) score≥4 scheduled for elective surgery were randomly divided into 2 groups(n=80 each):control group and study group.In control group the anesthesiologists visited their patients the day before operation ns usual;while in study group the anesthesiologists brought their patients to the operating room,showed them the environment and anesthesia equipment and assured of the safety of the operation and anesthesia.The degree of fear was scored according to FVAS (O=no fear,1-3 mild,4-6 moderate,7-10 severe) and was evaluated on the 1st morning after admission(T1),the night before operation(T2)and before induction of anesthesia(T3).MAP and HR at T1-3 and blood glucose level at T1,3 were mensured and recorded.Results The patient's preoperative apprehension was significantly allayed by patient's preoperative visit to the operating room.The MAP,HR,incidence of moderate and severe fear at T2,3 and blood glucose level at T3 were significantly lower in study group than in control group.Conclusion Patient's preoperative visit to the operating room can allay precperative apprehension.

The effect of plasma parathyroid hypertensive factor (PHF)from 153 essential hypertensive subjects on pressor response in normotensive rats was evaluated and plasma levels of parathyroid hormone (PTH) in these patients were measured. The results showed that PHF from 61 patients could induce a delayed increase in blood pressure in the normotensive rats. Mean arterial pressure (MAP) began to increase at 10 minutes after injection and a peak response was reached at 50 minutes while pre-injection MAP level was recovered at 70 minutes. On the other hand, plasma PHF in remaining 92 patients failed to induce a pressor effect. Plasma PTH in the 153 patients were considerably higher than those of the control, and the plasma PTH from the two groups, either with or without PHF, showed significant difference in comparison with the control, but plasma PTH between these two groups did not show significant difference. The results have suggested that pressor effect was independent of PTH but depends on PHF.

Objective:To investigate overall quality status of rose bengal agar culture medium for drug control in China to provide technical support for the drug standard revision. Methods:Four quality control microorganisms, namely Aspergillus niger, Candida al-bicans, Staphylococcus aureus and Escherichia coli, were used to evaluate the quality of rose bengal agar culture medium from 7 domes-tic and foreign manufacturers. Results:Including the control medium, the pH value of all the 8 media was different. The growth rate of Aspergillus niger and Candida albicans in the seven tested media were all above 0. 7 without significant difference (P>0. 05). The selectivity factors of Staphylococcus aureus in the seven media and the control medium was more than 6. The eight media showed differ-ences in the selectivity factors of Escherichia coli. Among them, A, B, C, E, G and the control medium had no inhibitory ( selectivi-ty) , while D and F showed certain selectivity. Conclusion:The rose bengal agar media from different manufacturers for drug control show similar growth-promoting ability with different pH value and selectivity to Escherichia coli.

BACKGROUND: It is proposed that elevated serum homocysteine is an important independent risk factor for ischemic stroke (IS), and 5, 10-methylenetetrahydrofolate reductase (MTHFR) is the key enzyme for homocysteine metabolism. The relationship between genetic mutation of MTHFR and IS remains controversial.OBJECTIVE: To examine the association of hyperhomocysteinemia and two MTHFR gene polymorphisms with IS in Northwest Chinese population.DESIGN: Case-control study.SETTING: Department of Neurology, First Hospital Affiliated to Jilin University, and Department of Neurology, Xijing Hospital, Fourth Military Medical University of Chinese PLA.PARTICIPANTS: Ninety-seven consecutive patients with ischemic stroke (71 males and 26 females) treated between November 2001 and May 2002were recruited, who were diagnosed by CT scan or MRI in the Department of Neurology, Xijing Hospital, Fourth Military Medical University of Chinese PLA. The control group consisted of 94 subjects (58 males and 36 females) without history of ischemic stroke. All the subjects were free of intracranial hemorrhage, cancer, renal dysfunction, and none used multivitamins or estrogen.METHODS: Serum homocysteine was measured by fluorescence polarization immunoassay. Polymerase chain reaction-restriction length polymorphism (PCR-RFLP) method was employed to detect the genotype at the two sites of C677T and A1298C in MTHFR gene.MAIN OUTCOME MEASURES: Serum homocysteine levels and the genotypic frequency frequencies of the two mutations of MTHFR.RESULTS: The 677T allele frequency was 59.3% in IS patients and 44.7% in the controls, showing significant differences (P=0.006), but no difference in 1298C allele frequency was detected between the two groups (22.7% vs 19.7%, P ＞ 0.05). Homozygous 677TT genotype was closely associated with hyperhomocysteinemie (P ＜ 0.01). In multivariate logistic regression analysis,677T gene mutation and hyperhomocysteinemie were all associated with the IS, with an OR of 1.870 and 1.031 (P＜ 0.05), respectively.CONCLUSION: Hyperhomocysteinemie is a risk factor of IS, and C677T mutation significantly increases homocysteine levels, and serves also as an independent genetic risk factor of IS.

Objective To explore a new method of establishing rat model of acute myocardial infarction by off ventilator support,spurt and repeated cryoinjury.Methods Sixty Wistar rats with male and female in half were randomized into three experimental groups according to cryoinjury time with 20 rats in each group:15-second group(cryoinjury for 5 s each time,three times in total),25-second group(5 s each time,five times in total)and 40-second group(5 s each time,eight times in total).The heart was exposed through a 1.0-to 1.5-cm left lateral thoracotomy.Cryoinjury of the LV free wall(LVFW)was performed by applying for 5 s a round copper probe of 6-mm in diameter immersed in liquid nitrogen for 5 min to reach-190 ℃,then the thoracic cavity was closed immediately.When the rats' breath restored,the cryoinjury of the LVFW in situ was repeatedly to ensure a transmural injury according to the protocol.The survival rate during operation and in 28 days after operation was observed.Also infarct size was observed by heart pathological section stained with Masson's trichrome on day 28 after operation.Results Infarct size in all three groups was larger than 20% signified by transmural injury,and completely myocardiolysis and distinct infarct boundary were observed.The survival rate was 16/20 in 15-second group,5/20 in 40-second group(P0.05 vs 15-second group and 40-second group).The infarct size in 40-second group was larger than the other two groups(standard deviation:10.564 4% vs 5.192 6%,6.496 0% in 15,25-second group,respectively)and its survival rate was lowest.The procedure of 25-second group was more complicated and had a tendency of higher death rate than that in 15-second group.Conclusion Using spurt,repeated ultra low temperature cryoinjury for 15 s may be an ideal method to establish the rat model of acute myocardial infarction.

Objective:To provide the basic data for the further revision of mycoplasma test method described in Chinese Pharma-copoeia and some references for the operation standardization of drug mycoplasma test. Methods:Two incubation conditions,namely aerobic conditions and microaerophilic conditions,were compared with respect to the growth status of mycoplasma in liquid and solid media. Results:The growth of mycoplasma was obvious difference between the two incubation conditions,and the microaerophilic con-ditions were better than the aerobic conditions. Conclusion:The microaerophilic conditions can be used in the incubation of mycoplas-ma test,which should be defined and standardized in the future Chinese Pharmacopoeia.

Microbiological data deviations ( MDD) investigation differs from out of the standard ( OOS) investigation in chemical inspection items. Investigating Out-of-Specification( OOS) Test Results for Pharmaceutical Production issued by FDA specifically ex-cludes microbiological and other biological assays. Until recently, there is not a definitive guideline for MDD investigation in the world. Some institutions and experts have explored the practice and theory of MDD investigation. Referred to the existing practical experience and theoretical achievement and based on the provisions of Chinese GMP (2010 revision) and the requirements of Chinese Pharmaco-poeia Ⅱ(2010 edition) Appendix XIX Q, the article discussed the methods for MDD laboratory investigation.

Objective:To investigate the growth-promoting properties and applicability of the mycoplasma test culture media pre-scribed in Chinese pharmacopoeia, and provide reference for the standardization of the drug mycoplasma test method. Methods: Re-spectively using four quantitative detection methods including sensitivity, degree of color change, colony counts and colony diameter, and mycoplasma media widely used in the world as the reference media, the growth-promoting properties of 4 batches of mycoplasma broth and 4 batches of arginine mycoplasma broth from four domestic manufacturers were investigated. Results:The results of sensitivity assay and absorbance detection showed that all the media inoculated with below 100 CFU test microorganisms exhibited visible color change. Furthermore, the results of color change degree and colony diameter showed that there were significant differences among the media products from different manufacturers(P<0. 01). Conclusion:Mycoplasma broth and arginine mycoplasma broth both can sup-port the growth of below 100 CFU test microorganisms. Due to the difference in the growth-promoting properties among the media prod-ucts from different manufacturers, the drug mycoplasma test workers should use more sensitive methods to examine the applicability of the media.

BACKGROUND:Many liver cancer stem cel markers have been found in liver cancer tissues and cel lines such as CD133, acetaldehyde dehydrogenase (ALDH), CD90, CD44, EpcAM, CD13, OV6, K19, c-kit and ABCG2. Of them, CD133, CD90 and CD44 have been shown to be strongly associated with the recurrence and metastasis of liver cancer. OBJECTIVE:To explore the dynamic changes of liver cancer stem cel markers and inflammatory factors during the induction of liver cancer in rats and their correlation. METHODS:Diethyl nitrosamine solution was given to Sprague-Dawley rats for 24 hours to induce rat models of liver cancer. Rats that were given common water were considered as the healthy control group. RESULTS AND CONCLUSION: Immunohistochemical staining revealed that Kupffer cels-related ED2 expression showed a gradual increase in the model group. Compared with the healthy control group, ED2 expression was significantly higher at 12, 16, 20 and 24 weeks after induction in the model group (P < 0.05). Quantitative PCR demonstrated that CD90 showed a gradualy increased trend during induction (P < 0.05). Compared with healthy tissue, CD90 increased significantly in the liver cancer tissue (P < 0.05). CD133 showed an increased trend, but one-way analysis of variance did not show significant differences (P > 0.05). During induction, no significant change was found in other liver cancer stem cel markers (P> 0.05). During the induction, tumor necrosis factor α, transforming growth factor β, MCP-1 and interleukin-6 expression levels were significantly increased (P < 0.05). Compared with healthy tissue, transforming growth factor β, MCP-1 and interleukin-6 expression levels were significantly higher in the liver cancer tissue (P < 0.05). Other inflammatory factors did not exhibit significant alterations during the induction (P > 0.05). Pearson correlation analysis demonstrated that MCP-1, transforming growth factor βand interleukin-6 expression levels were significantly positively correlated with CD90 expression (P < 0.05). These findings suggest that partial inflammatory factors released from Kupffer cels have a certain correlation with liver cancer stem cels. Kupffer cels can promote the occurrence of liver cancer.