Objective To find a preferable cryopreservative solution and enhance the viability further.Methods Donor hepatocytes were isolated from hybrid pig weighting 5～8 kg by means of collagenase.Then the hepatocyte suspension was put into a liquid nitrogen container for one month with three different cryoprotectants (only use DMSO,combined use of DMSO+HES and combined use of DMSO+HES+1.3 mmol/L Ca 2+).The samples were thawed rapidly and detected in the viability and morphology after a month.Fractional hepatocytes were cultured five days in containing Ca 2+ and uncontaining Ca 2+ culture media to observe the expression of function.Results Porcine hepatocytes,which were preserved in different culture media,show the different viability and morphology manifestation.Among the total the method of only uses DMSO show worst effect.Then is the DMSO+HES group.Combined cryopreservative method with HES+DMSO+1.3 mmol/LCacl 2 has a best effect and functional expression.Hepatocytes were cultured in culture media containing Ca 2+is better than those were cultured in culture media uncontaining Ca 2+.Conclusion The combined cryopreservative system in which HES+DMSO+1.3 mmol/LCacl 2 as the cardinal component can improve the preservative quality significantly.

Objective To investigate the safety and clinical significance of total thyroidectomy performed for benign thyroid disease. Methods Eighty-eight patients with benign thyroid disease were treated with total thyroidectomy. The postoperative complications were analyzed. Results With primary total thyroidectomy, the incidences of transient hypocalcemia and transient recurrent laryngeal nerve paralysis were 2.5% and 1.2% respectively. The incidences after reoperation were 28.6%( P

Objective To study the value of the neuron-specific enolase(NSE) in the serum of patients with cerebral stroke in diagnosis, seriousness and evaluation of prognosis. Methods The levels of NSE in the serum of 77 patients with cerebral stroke(36 cases with hemorrhage, 41 cases with infarction) were measured by radio-immunoassay (RIA)method.Results The serum leves of NSE both in cerebral infarction and cerebral hemorrhage patients were obviously higher than those of the control group( P 0.05). In the first week there was a positive correlation between NSE serum level and neurological deficit scores,the volume of haemorrhage, the volume of infarction( P 0.05) . Conclusion The NSE level in serum may be served as a objective index of seriousness and early diagnosis in stroke, but no helpful in differential diagnosis and evaluation of prognosis.

Huoxue Tongmai Lishui method, a traditional Chinese medicine treatment for eliminating water, activating and promoting blood circulation, could inhibit fundus hemorrhage on experimental retinal vein occlusion (RVO) with high obvious effective rate, and improve symptoms in traditional Chinese medicine. The action mechanism may be related to reducing plasma viscosity and non-perfusion area, and the formation of collateral circulation.

OBJECTIVE To study the distributive properties and susceptibility of yeasts to six antifungal agents. METHODS To analyze the distributive properties of 264 clinical Candida spp isolates and study the susceptibility to amphotericin B,nystatin,fluconazole,ketoconazole,miconazole and clotrimazole.The susceptibility of yeasts was tested according to the National Committee for Clinical Laboratory Standards guideline(NCCLS M27-A2). RESULTS Strains of Candida albicans were the most frequent organism isolated accounted for 62.5% of all the isolates.C.tropicalis,C.glabrata,and C.parapsilosis accounted for 20.8%,12.5%,and 1.9%,the others accounted for only 2.3%.The main infected organs were lungs,urinary tract,and digestive tract;the susceptibility tests showed strains of Candida spp to nystatin,amphotericin B,and fluconazole were more active than to the other antifungal agents.The resistance to triazole antifugal agents could be shown. CONCLUSIONS We should strengthen the diagnosis of Candida spp and strengthen the surveillance on susceptibility of clinical isolates Candida spp so as to help the doctors choose the antifungal agents reasonably.

AIM To construct a non-normalized cDNA library from Agkistrodon acutus venom gland as an imtial step to develop new and more effective venom by genetic engineering technique for screening and expressing target genes. METHODS The total RNA was extracted from fresh venom gland using Trizol. mRNA was reversely transcripted to cDNA using superscriptⅡ reverse transcriptase. Second-strand synthesis was performed using DNA polymeraseⅠ. After adding EcoRⅠ adaptor, phosphorylating the end and digesting with XhoⅠ, the cDNA was collected in five fractions (<0.25 kb, 0.25-0.5 kb, 0.5-1 kb, 1-2 kb and >2 kb) using the QIAquick Gel Extraction kit and ligated to pBluescriptⅡ vectors. The five libraries obtained were plated by infecting E.coli DH10B, constructing a cDNA library of Agkistrodon acutus venom gland. Sequencing clones at random, 8696 high quality 5′ end expressed sequenced tags (ESTs) were obtained and analyzed. The initial sequences were assembled into 2855 clusters. Among which, one of the clusters (Agkihagin) consisting of 74 ESTs was identified as a novel metalloprtoteinase based on RT-PCR and sequence analysis. RESULTSThe titers of library were 2.048×106. The novel metalloproteinase belonged to PⅢ type metalloproteinase. Its open reading frame was composed of 1827 nucleotides and coded a pre-zymogen of 608 amino acid with zinc-binding domain for metalloproteinase and Asp-Glu-Cys-Asp(DECD) domain for disintegrin. CONCLUSION The capacity of cDNA library of venom gland is above the general level of cDNA library. It would be a helpful platform to construct a catalog for transcripts in the venom gland of the Agkistrodon acutus. The sequence analysis indicates that the deduced amino acid sequence of the identified gene for metalloproteinase share the highest 87% identity with the metalloproteinase genes of other snakes in the GenBank. It lays a good foundation for the study of structure-function relationships of snake venom metalloproteinases.

Objective To evaluate the blood-saving effect of tranexamic acid in patients undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB).Methods The study was a prospective,randomized and placebo-control trial.Two hundred ASA Ⅰ-Ⅳ patients,aged 18-64 yr,weighing 50-100 kg,were randomized to receive placebo (group C,n =100) or tranexamic acid (group T,n =100).Tranexamic acid 10 mg/kg was intravenously infused over 20 min before skin incision followed by continuous infusion at 10 mg· kg-1 · h-1 until the end of operation in group T.While the equal volume of normal saline was given in group C.The total volume of postoperative chest tube drainage,postoperative massive bleeding and a second thoracotomy for stopping the bleeding were reordered.The requirement for transfusion of allogeneic blood and complications during the perioperative period were also recorded.Results Compared with group C,the total volume of postoperative chest tube drainage and incidences of postoperative massive bleeding and a second thoracotomy for stopping the bleeding were significantly decreased,and the requirement for transfusion of allogeneic red blood cells,platelet and fresh frozen plasma was reduced in group D (P ＜ 0.05).There was no significant difference in the incidence of complications between the two groups (P ＜ 0.05).Conclusion Tranexamic acid exerts the blood-saving effect in patients undergoing CABG with CPB and can significantly reduce postoperative bleeding and transfusion of allogeneic blood.

BACKGROUND: Our previous study showed that the activation of local renin-angiotensin system in heart and vessels contributed to hypertension and cardiovascular remodeling. However, whether oxygen free radical plays an important role in this process is still unclear. OBJECTIVE: To investigate the interventional effects of Ebselen, a kind of anti-oxidative drug, on rats administered by Nw-Nitro-L-arginine methyl easter (L-NAME) (L-NAME), inhibitor of nitric oxide synthase (NOS) for a long term, and probe into the role of oxygen free radicals (OFR) in hyper- tension and cardiovascular remodeling of NO-deficient rats. DESIGN: A randomized grouping and controlled animal trial. SETTING: Department of Cardiology, the Second Affiliated Hospital of Nanchang University, Institute of Cardiology, Nanchang University. MATERIALS: The experiment was completed from January 2002 to March 2003 at the Animal Experimental Lab, Institute of Cardiology, Nanchang University and the Key Molecular Medical Lab of Jiangxi Province. Twenty-four Wistar rats were divided to three groups according to the random number table method: normal control group (n=8), L-NAME group (n=8), L-NAME + Ebselen group (n=8). METHODS: ①Normal control group: The rats could eat and drink routinely, and they were administrated by skim milk ball (net weigh = 4 g) before feed every night. ② L-NAME group: The rats received L-NAME in the dose of 50 mg/kg mixed in one skim milk ball everyday before feed every night. ③ L-NAME + Ebselen group: The rats were admin istered by one skim milk ball (net weigh = 2 g) mixed with L-NAME (50 mg/kg) and one skim milk ball (net weigh = 2 g) mixed with ebselen (30 mg/kg). The systolic blood pressure (SBP) was detected before LNAME was given and at the ends of the 1st, 2nd, 4th,6th and 8th weeks respectively. The rats were killed under anesthesia at the end of the 8th week, the plasma and homogenate of myocardium of apex were taken to detect the biochemical indexes, the other heart tissues were used for the histological detections. MAIN OUTCOME MEASURE: ① Dynamic changes of blood pressure were observed. The levels of NO and malondialdehyde (MDA), activities of angiotensin-converting enzyme (ACE) and superoxide dismutase (SOD) in plasma and myocardium of apex were measured. The production of superoxide anion and the levels of angiotensin Ⅱ type 1 receptor (AT1R) protein expression in myocardium of apex were determined. ③ The pathomor- phological indexes were determined. RESULTS: All the 24 rats were involved in the analysis of results without deletion. ① SBP: At the ends of the 1st, 2nd, 4th 6th and 8th weeks, SBP elevated gradually in the L-NAME group, which were significantly higher than those in the control group at the same time (P ＜ 0.05-0.01). At the end of the 8th week, the SBP was significantly lower in the L- NAME + Ebselen group than in the L-NAME group (P ＜ 0.05). ② Bio chemical indexes in plasma and homogenate of myocardium of apex: The NO level and SOD activity in myocardial tissue were significantly lower in the L-NAME group than in the normal control group (P ＜ 0.05-0.01), but significantly higher in the L-NAME + Ebselen group than in the L- NAME group (P ＜ 0.05-0.01). The production of superoxide anion, ACE activity and level of AT1R expression were all significantly higher in the L-NAME group than in the normal control group (P ＜ 0.05), but signifi- cantly lower in the L-NAME + Ebselen group than in the L-NAME group. ③ Pathomorphological indexes: The ratio of cardiac mass to body mass, thickness of left ventricle and ratio of thickness to lumen diameter of small artery were all significantly higher in the L-NAME group and L- NAME + Ebselen group than in the normal control group (P ＜ 0.05-0.01), and the thickness of left ventricle and ratio of thickness to lumen diameter of small artery were significantly lower in the L-NAME + Ebselen group than in the L-NAME group (P ＜ 0.05) CONCLUSION: Ebselen, an anti-oxidative drug, enable to attenuate the development of hypertension and cardiovascular remodeling in NO-deficient rats. By activating renin-angiotensin system (RAS), OFR may accelerate the formation of hypertension and cardiovascular remodeling, and the increased production of OFR may contribute to the development of cardiovascular remodeling. It is indicated that RAS may play an important role in the development of hypertension and cardiovascular remodeling induced by NO-deficiency

OBJECTIVE:To establish the system of curriculum based on working process and occupation ability for pharmacy major in higher vocational colleges. METHODS:Investigation was conducted among medical practitioners from pharmaceutical companies,hospitals,pharmaceutical factories,scientific research institutions and other related professionals. RESULTS:150 ques-tionnaires were sent out,and 141 valid questionnaires were collected with effective recovery rate of 94.00%. Results of investiga-tion showed that respondents most valued graduates with interpersonal and communication skills,followed by professional skills and practical ability. They were mainly clinical application of drugs,pharmacological effects and adverse reactions of drugs in the pharmacology theory teaching,the mechanism of action of drugs were weakened. The ability of prescription distribution,symptoms inquiring and drugs recommending should be strengthened in the pharmacology theory practice teaching. More than half of the re-spondents thought that confirmatory tests were necessary to keep,which helped to train students’practical ability and deepen the understanding of the theory. Meanwhile,it was important to strengthen the students’communication with the patients or their fami-lies and doctors to cultivate the ability of acquiring professional knowledge. CONCLUSIONS:The investigation provides basis for the making of curriculum standards of pharmacology,through which teaching contents are selected,teaching methods are de-signed,and it makes the pharmacology course full of post applicability and provides better decision-making basis to meet the posi-tion requirements.

Objective To study the expression of leukemia-related protein 16 (LRP16) in human pituitary prolactin adenomas (PRL adenomas), and the relationship between LRP16 and estrogen receptorα (ERα).Methods From October 2009 to September 2014, thirty-one adult patients diagnosed and pathologically confirmed as pituitary prolactin (PRL) adenomas (observation group) and 22 pituitary non-PRL adenomas (control group) in the Chinese PLA General Hospital were verified by the pathological examination. Immunohistochemical method was used to detect the expression levels of LRP16, PRL and ERα.Results Of the patients in observation group, those aged under 30 were predominantly females, while male patients were more common in those aged over 30. LRP16 positively existed in 26 cases (83.9%, 26/31), and ERα was positive in 28 cases (90.3%, 28/31), the both ratios were significantly higher than those in control group. ERα and LRP16 increased synchronously both in expressive content and intensity, showing a certain positive correlation between them.Conclusions Higher body mass index may be a high-risk factor badly affected the occurrence, development and prognosis in male patients with pituitary PRL; LRP16 may take part in the proliferation and formation of pituitary PRL adenomas through ERα.