TMP. The results suggested that 12D-TMP and 6D-TMP possessed remarkable isotope effects, and that the pyrazine ring of TMP may be responsible for its pharmacodynamics, while the substituted groups might primarily govern its pharma-cokinetics and toxicity. The results also indicated that to study the structure activity relationship of TMP and to modify its chemical structure are very fruitful for the development of TMP analogues of high potency.

OBJECTIVE　To determine the pharmacokinetic parameters of perlolyrine in rats.METHODS　The plasma concentration and pharmacokinetic parameters of perlolyrine were determined by means of GC-MS with selected ion (m/z 247 and m/z 248) and ［2-15N］perlolyrine(m/z 248) as the internal standard.RESULTS　The concentration-time profile of perlolyrine after oral administration of perlolyrine fitted a two-compartment open model.The pharmacokinetic parameters were t1/2α=0.31 h, t1/2β=4.62 h, t1/2ka=0.10 h, tmax=0.34 h, cmax=18.74 ng.mL-1,K12=0.82 h-1, K21=0.38h-1,K10=0.29 h-1,Vb=108.16 L.kg-1,AUC=98.54 ng.h.mL-1,respectively.CONCLUSION　The method is constant,sensitive and accurate.It provided a scientific basis for the clinical use of perlolyrine.

Experimental research showed that Gan Tang Zhi can influence blood clotting and plasma fibrinolytic system of rabbits, 15min after introvenous injecting 6.25mg/kg or 25mg/kg Gan Tang Zhi into rabbit it can pronouncedly prolong prothrombin time ( PT ) , thrombin time ( TT ) , and kaolin partial thromboplastin time ( KPTT ) ; and cause remarkable increase of fibrin degradation product ( FDP ) , Gan Tang Zhi can also shorten englobulin lysis time (ELT) and decrease fibri-nogen.

The influences of propylene glycol mannate sulfate (PGMS) on experimental thrombosis and thrombolysis in vivo were studies after iv 8. 125, 6. 25, 12. 5 , 25mg/Kg in rabbits, and the effect of antithrombosis of PGMS was compared with that of heparin. The results showed that PGMS possessed remarkable effect of antithrombosis. In order to explore the mechanism of antithrombosis of PGMS, we studied the influences on the fibrinolytic and coagulant function of rabbits. The results showed that PGMS can pronouncedly prolong the prothrombin time (PT), thrombin time (TT), kaolin partial thromboplastin time (KPTT), and enhance the activity of antithrombin-III (AT -III). PGMS can cause a remarkable increase in fibrin degradation product (FDP) , shorten euglobulin lysis time (ELT) , and a decrease in the contents of fibrinogen and plasminogen activity. These results suggested that PGMS probably exert the antithrombotic effect by inhibiting coagulation and activating fibrinolysis.

ObjectiveTo explore the efficacy of sevoflurane or propofol combined with remifentanil during the maintenance of general anesthesia in thyroid gland surgery.MethodsSixty patients underwent thyroid gland surgery were randomly divided into tow groups.Once the larynx mask was intubated, anesthesia was maintained with propofol(the effect site concentration was 2.5 ～3.5mg/L) and remifentanil(the effect site concentration was 4.5 ～5.5μg/L) by TCI technique in group P,but with sevoflurane(2％～4％) and remfentanil(the effect site concentration was 2.5 ～4.0g/L)in group S.The depth of anesthesia was measured by the A-line TM AEP Monitor which expressed as A-Line ARX Index TM(AAI).All patients' SBP,DBP and HR were recorded at eight time points: before induction time(T0) ,after induction but before larynx mask intubation time(T1) ,intubate larynx mask time(T2) ,cut skin time (T3), separate thyroid gland time (T4), cut thyroid gland time (T5), remove larynx mask time (T6) ,leave the operation room time(T7).The emergency time,the conscious of the patients after anesthesia and the side effects were also recorded.ResultsThere were no significant differences between the groups with respect to age,gender,weight,the duration of operation,the emergency time and the conscious of the patients after anesthesia.SBP,DBP,HR of the patients in both groups showed no significant difference at T0,T1 ,T2, T3 (all P ＞ 0.05), but had significant difference at T4,T5,T6, T7 (all P ＜ 0.05).Compare with group P,the incidentce of restlessness, dizziness, drowsiness, rigor and pain was significantly lower in group S(all P ＜0.05).The incidentce of nausea,vomit and aspiration did not appear in both groups.ConclusionBoth groups showed good anesthesia effects and the patients also emerged from anesthesia quickly.But the anesthesia maintained with sevoflurane and remifentanil could bring more stable hemodynamics and lower incidence rate of the side effects.

Objective To evaluate the safety and efficacy of endoscopic treatment in patients with primary hepatic carcinoma and gastroesophageal variceal hemorrhage. Methods Thirty cases of hepatic carcinoma with gastroesophageal variceal hemorrhage diagnosed with gastroscopy between January 2008 and June 2015 in Chinese PLA General Hospital and Hainan branch were included. Patients′ clinical data, complications and therapeutic effects of endoscopy were retrospectively analyzed. Results All patients underwent endoscopy successfully. No recurrent bleeding was found in any case after treatment. Eight cases of varicosis nearly disappeared. Six?month follow?up showed three cases of gastrointestinal bleeding, one case of death of bleeding, and no recurrence hemorrhage in 27 others. Conclusion Endoscopic treatment could safely and effectively control the primary liver cancer combined with gastroesophageal variceal hemorrhage, hence may provide opportunity for further treatment of liver cancer.

AIM: To obtain the glycopohorin A (GPA) cDNA and construct the target gene in yeast two-hybrid.METHODS: About 410 bp cDNA fragment was amplified from K562 cell by RT-PCR.After being sequenced, the GPA gene fragment was cloned into EcoR -Ⅰ- Pst Ⅰ site of pbridge to form BD ends in yeast two-hybrid system. The recombinant plasmid was transfered into yeast AH109, and the expression in the yeast was also examined. RESULTS: The amino acid sequence encoded by cloned cDNA was mostly the same as reported GPA, and about 1 mm white yeast clone grew in the selective medium after 3 d.CONCLUSION: pbridge-GPA has nontoxic to the yeast, which can serve as a target gene in yeast two-hybrid system.

AIM: To study the effect of the overexpression of p16 on an anion exchange function of band 3 in HeLa cells. METHODS: The expression of p16 and band 3 in HeLa cells was detected by immunohistochemistry (IHC). The p16 cDNA was subcloned to plasmids pEGFP-C1 by PCR and identified by restriction enzyme digestion and sequencing, and then, the recombinant pEGFP-C1-p16 plasmids were transiently transfected into HeLa cells. The expression of fusion protein in HeLa cells was detected by fluorescence microscope. 6-methoxy-N-(3-sulfopropyl)-quinolinium(SPQ)fluorescent probes were used to detect the anion exchange function of band 3. RESULTS: P16 and band 3 were expressed in HeLa cells. The amplificated p16 cDNA sequence was the same as the report sequence. The transfective efficacy of pEGFP-C1-p16 was above 60%. The anion exchange function increased after the transfection of pEGFP-C1-p16 plasmids. CONCLUSION: p16 facilitates the anion exchange function of band 3 in HeLa cells.

[Objects]To isolate and identify the pathogen of the large outbreak of dengue in Guangdong province in 2014. To understand the origin and the phylogenetic characteristics of the isolates ,and provide scientific foundation for the surveillance and prevention of dengue fever.[Methods]Collected the patient serum samples over all the Guangdong province during the 2014 outbreakperiod,isolated and identified the virus from these samples. Amplified complete E gene and complete genome with certain primers and sequenced all the products. Then the Phylogenetic ,Bayesian phylogeography and mutations analysis were carried.[Results]40 DENV-1 strains were isolated and identified. 40 complete E gene sequences and 6 complete genome sequences of DENV-1 were obtained. Phylogenetic analysis with E gene sequences revealed that the 40 isolates were classified into two genotypes including 16 genotypeⅠ(Asia)and 24 genotypeⅤ(America/Africa). 14 genotypeⅠisolates were clustered closest with isolates from Guangdong province(2013)and Sigapore(2013)which share the nucletide identities of 99.6% ~ 99.9%,other two genotypeⅠisolates were clustered with strains from Malaysia (2013) and both share the nucletide identities of 99.7%;24 genotypeⅤisolates were all classified in one clade with striains from Bangladesh(2009),China(2009)and Bhutan(2013)which share nucletide identities of 99.0%-99.9%. Further analysis with six complete genome sequences showed that five isolates were clustered closest with strains isolated from Guangdong province(2013)share the nucletide identities of 99.6%-99.8% while the sixth stains closest with strains isolated from Myanmar(2002)share the nucletide identities of 98.8%. The isolates have five amino acid mutations compared with strains epidemic in Guangdong province in 2013,three mutations(S88V,E203G,T275R)are in the EⅡdomain and one mutation (S305P)is in the EⅢdomain which associated with virulence.[Conclusions]During the outbreak in Guangdong province in 2014, DENV-1 is the predominant causative serotype,and there are at least two different kinds of genotypes of DENV-1 largely epidemiced in the whole province. Evolution analysis reveals the multiple origins of the isolates which may origin from Guangdong province , Sigapore,Malaysia,Myanmar so that we should enhance the study and surveillance of autochthonous and vectors in order to understand the epidemic way of dengue in Guangdong province. The isolates have had four mutations in the domain associated with virulence which remain further study to know their biological effects.