Glycyrrhiza uralensis Fisch. ex DC is widely used in traditional Chinese medicine (TCM). Among its various active components, glycyrrhizic acid is believed to be the marker component. Squalene synthase (SQS) and beta-amyrin synthase (beta-AS) are key enzymes in the biosynthetic pathway of glycyrrhizic acid in G uralensis. To reveal the effects of co-expression of SQS1 and beta-AS genes on this pathway, 7 yeast expression vectors harboring different SQS1 variants and beta-AS were constructed and expressed in Saccharomyces cerevisiae as fusion proteins. TLC and GC-MS results showed that co-expression of SQS1 and beta-AS enhanced the accumulation of beta-amyrin. The effects of SQS12 were more obvious than the other two SQS1 variants. This study is significant for further investigations concerned with exploring the biosynthesis of glycyrrhizic acid in vitro and strengthening the efficacy of G. uralensis by means of increasing the content of glycyrrhizic acid.
Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Glycyrrhiza uralensis ; genetics ; Intramolecular Transferases ; metabolism ; Oleanolic Acid ; analogs & derivatives ; metabolism ; Plant Proteins ; genetics ; Recombinant Proteins ; metabolism ; Saccharomyces cerevisiae ; metabolism

On the basis of the Uncaria transcriptome, specific primers were designed for UrSTR. The full-length cDNA of UrSTR (GeneBank: OL310251) was 1 541 bp, encoding 345 amino acid residues, and the promoter region sequence of UrSTR (GeneBank: OL310252) was 1 179 bp. Phylogenetic tree is revealed that UrSTR had a closest relationship with STR from Ophiorrhiza pumila and Ophiorrhiza japonica. Localization of UrSTR protein is revealed located in the vacuole membrane. Plant-care analysis indicated that the promoter region sequence of UrSTR, covering multiple light, stress and hormone-response cis-regulatory elements, and verified transcriptional activity. The results of SDS-PAGE show that pET-28a-UrSTR recombinant protein was successfully expressed, and the size was anticipated. The UrSTR prokaryotic expression system needs to be optimized in the later stage. The research lays the foundation for further purification to study its structure and functional characterization of the UrSTR protein.

Objective To analyze the efficacy of different treatment methods for end-stage hepatic alveolar echinococcosis combined with obstructive jaundice.Methods A retrospective study was conducted on the diagnosis and treatment of 55 patients with end-stage hepatic alveolar echinococcosis combined with obstructive jaundice managed from January 2000 to January 2013 at the First Affiliated Hospital of Xinjiang Medical University.The patients were divided into two groups according to the treatment options:group A,the palliative surgery group (n =38,69.1％) using palliative resection and biliary decompression ; and group B,the interventional group (n =17,30.9％) using percutaneous transhepatic cholangial drainage (PTCD).We analyzd the general data,preoperative and postoperative liver function,operation time,blood loss,average hospital stay,duration of postoperative tube drainage of abscess cavity,degree of lesion with invasion into the first porta hepatis,progressive lesion,continuous invasion and/or distant metastasis,biliary complications,mortality,and cumulative survival rates.The t-test or t'-test was used to analyze continuous data and the chi-square test was used to analyze categorical data.Parallel log rank test and Kaplan-meier method were used to calculate survival rates in survival analysis.Results When compared with group B,group A had significantly longer operative time,more blood loss,and longer average hospital stay (P ＜0.05).The postoperative total bilirubin,direct bilirubin,γ-glutamyl transpeptidase and alkaline phosphatase,aspartate aminotransferase,alanine aminotransferase were significantly higher than in group B (P ＜0.05).The level of post operative albumin was lower in group A than in group B.The two groups of patients (A and B) had similar background including general data (gender,age,nation),preoperative liver function,duration of postoperative tube-drainage of abscess cavity,degree of lesion-invasion into first porta hepa tis,progressive lesion-invasion and/or distant metastasis,biliary complications,mortality,cumulative survival rates and survival curves.Conclusions Interventional treatment is an effective treatment for end-stage hepatic alveolar echinococcosis combined with obstructive jaundice.It has the advantages of minimal invasiveness,simplicity,safety and repeatability.It may replace traditional palliative surgery in the future.

Objective To analyze the use of multidisciplinary team approach (MDT) for advanced and refractory hepatic alveolar echinococcosis in individualized treatment.Methods A retrospective study was conducted on the use of multidisciplinary team approach in individualized diagnosis and treatment for 137 patients with advanced and refractory hepatic alveolar echinococcosis (with invasion of major blood vessels and bile ducts,and/or with lung and brain metastasis) in our hospital from January 2005 to December 2013.The patients were divided into two groups:The MDT group (n =49) and the non-MDT group (n =88).The MDT group was further divided into two subgroups:subgroup A was the surgical treatment group (n =26),and subgroup B was the non-surgical treatment group (n =26).In the subgroup B,13 patients underwent late radical surgery.The non-MDT group was also further divided into two subgroups:subgroup a was the surgical treatment group (n =61),and subgroup b was the non-surgical treatment group (n =27).In subgroup b,5 patients underwent late radical surgery.The time taken to confirm the diagnosis,perioperative hospital stay,operation time,blood loss,postoperative drainage time,postoperative hospital stay,early postoperative complications (pleural and peritoneal effusions,bile leak,anastomotic leak),late postoperative complications (jaundice,anastomotic stenosis,recurrence),and rates of radical surgery were compared between subgroup A and subgroup a.The rates of late radical resection were compared between subgroup B with subgroup b.All data were analyzed using the Mann-Whitney rank sum test or the Chi-square test.Results Subgroup A had significantly shorter perioperative hospital stay,postoperative hospital stay,and total length of hospital stay than subgroup a (P ＜ 0.05).The incidence of late postoperative complications (jaundice,anastomotic stenosis,recurrence) was also significantly less than subgroup a (P ＜ 0.05),and the radical surgery rate was significantly higher than subgroup a (P ＜ 0.05).There was no significant difference in the time taken to confirm the diagnosis,operation time,blood loss,postoperative drainage time,early postoperative complications (pleural and peritoneal effusions,bile leak,anastomotic leak) (P ＜ 0.05) between subgroup A and subgroup a.The ratio of subgroup B receiving chemotherapy alone or drainage + chemotherapy,and the rate of late implement of radical resection were significantly higher than subgroup b (P ＜0.05).Conclusions A multidisciplinary team approach in individualized treatment comprehensively combined the advantages of the effects of drugs,intervention,surgery and systemic nutritional support.The best individualized treatment plan could be used which improved the rates of radical surgery in advanced and refractory hepatic alveolar echinococcosis,reduced postoperative complications,improved quality of life,and offered chances of radical resection in the patients who had lost the opportunities for surgery.

Objective To evaluate the effectiveness and safety of radical versus conservative surgery for hydatid liver cysts.Methods The Pubmed, EMBASE, MEDLINE, Cochrane Library, Chinese Biomedicine Database, CNKI, Wan Fang Databases, VIP Databases, Chinese scientific and technical journals full-text database and Chinese Journal of full text database were searched for randomized clinical trials or none-randomized clinical trials from January 2000 to January 2015.A Meta-analysis was performed with RevMan5.1.Results 10 studies with 2 123 patients were included into the final analysis.Meta-analysis demonstrated that the radical group was significantly longer in operation time [weighted mean difference (WMD) =25.11;95％ CI: 16.18-34.05;P ＜ 0.05], was significantly shorter in the length of hospital stay (WMD =-3.94;95％CI:-7.57-0.32;P ＜0.05), was significantly lower in the incidence of bile leak (OR =0.19;95％ CI: 0.07-0.49;P ＜ 0.05), was significantly lower in the incidence of residual cavity infection (OR =0.08;95 ％ CI: 0.04-0.15;P ＜ 0.05), and in the incidence of local recurrence (OR =0.10;95 ％ CI: 0.05-0.18;P ＜ 0.05) when compared with the conservative surgery group.Conclusions For hydatid liver cysts, radical surgical procedures had significantly lower rates of complications and recurrence.Thus, they are better treatments for hepatic cystic echinococcosis.

Objective To explore whether human umbilical venous endothelial cells could be used as feeder layer to support the growth of embryonic stem cells (ESC) and keep ESC undifferentiated.Methods The venous vessels of umbilical cord obtained from healthy puerperal were perfused with collagenase.The isolated endothelial cells went through primary culture and passages for expansion.Factor Ⅷ antigens determination was implemented.Endothelial cells with good growth and 3 or above passages were treated with mitomycin-C(10 mg/L) and prepared as feeder layer,on which E14.1 ESC was transplanted for subculturing to observe the morphological characterization and determine ESC alkaline psphatase (AKP) activity and the expression of stem cell marker Oct-4.Severe combined immune deficiency(SCID) mouse in vivo terotoma formation experiment was performed to identify its pluripotent properties.Results Human umbilical vein-derived endothelial cells grew well in culturing in vitro and regenerate in large numbers.The endothelial cells maintained normal cellular morphological and biological characterization after 10 passages.The cells stopped proliferating after being treated with mitomycin-C,but its activity and morphological properties were well-maintained with 24 hours,which was a fundamental property of serving as feeder layer.E14.1 ESC remained undifferentiated in human umbilical venous endothelial cells after 3-8 passages,the cells grew in colony and showed high expression of AKP and stem cell Oct-4.In vivo pluripotency experiment showed that 6 weeks after being transplanted to SCID mice E14.1 ESC of 6 and 10 passages in endothelial cells both could form teratoma containing 3 layers of tissue cells.Conclusions Human umbilical venous endothelial cell serve as a convenient feeder layer cell with rich sources.It can effectively support ESC growth and heterogenous and prevent the heterogeneous protein pollution and pathogenic microorganisms caused by animal cell feeder layers,thus solve the problem of biological safety of ESC clinical application.

Objective To determine the effect of mild hypothermia on neonatal piglet cardiac hemodynamic function after hypoxia-ischemia (HI).Method Twenty five 7-day-old piglets were used for hypoxic ischemic brain damage (HIBD) model by the method of temporary occlusion of the bilateral carotid arteries and followed by mechanical ventilation with low concentration of oxygen (FiO_2=6%) for 30 minutes.The piglets were randomly divided into three groups:group A (normothermia with body temperature to 39℃,n=9),group B (body temperature to 36℃for 72 hours,n=8),and group C (body temperature to 34℃for 72 hours,n=8).Mild hypothermia was initiated at 4 hours after HI,the systolic and diastole function were evaluated by Doppler echocardiography at pre-HI,post-Hi 4 hours and post-HI 72 hours.Results There were no significant differences in left ventrieular ejection time/left ventrieular ejection time (LPEP/LVEF),right ventricular ejection acceleration time/right ventricular ejection time (RACT/RVET) and CO at post-HI with hypothermia 72 hours in three groups,but the heart rate decreased in B and group C group.Compared with nonnothermia,mild hypothermia treatment showed no significant differences in MAP,LPEP/LVET,RACT/RVET,CO,SV at post-HI with hypothermia 72 hours.Conclusions Body temperature decreased by 3～5℃for 72 hours will not aggravate hemodynamic abnormity.

Four new triterpenoids, together with six known analogues, were isolated from an aqueous extract of the Ziziphus jujuba var. spinosa seeds, by multiple column chromatographic separation methods using stationary phases of macroporous adsorption resin, MCI resin, normal phase silica gel, Sephadex LH-20, and Toyopearl HW-40C as well as preparative thin-layer chromatography and reversed-phase HPLC. Their structures were determined by spectroscopic data analysis, the new structures were trivially named jujubaceanothoside A (1), 23-epijujuboside A (2), and jujubosides J and K (3 and 4), while the known analogues were identified as jujubosides A-C (5-7) and II (8), alphitolic acid (9), and betulinic acid (10). The structure of 1 was confirmed by single crystal X-ray diffraction.

OBJECTIVETo establish a modified plasma protamine paracoagulation test.

METHODSPlasma protamine paracoagulation, modified plasma protamine paracoagulation and D-dimer (D-D) tests were performed for the plasma samples collected from 98 cases of disseminated intravascular coagulation (DIC) and 156 normal subjects. The sensitivity and specificity of the 3 tests were analyzed. The plasma samples from 8 cases of suspected myocardial infarction were detected using modified plasma protamine paracoagulation for diagnostic purpose.

RESULTSThe sensitivity of plasma protamine paracoagulation, modified plasma protamine paracoagulation and D-D tests was 16.33%, 88.76% and 77.56%, and the specificity was 100%, 88.46% and 97.44%, respectively. Positive results occurred earlier in modified plasma protamine paracoagulation test than in plasma protamine paracoagulation and D-D tests in 5 cases of myocardial infarction.

CONCLUSIONThe modified plasma protamine paracoagulation test has a higher sensitivity than plasma protamine paracoagulation test and a higher specificity than D-D test, and can be helpful in early diagnosis of thrombosis and fibrinolysis.

Adult ; Blood Coagulation Tests ; methods ; Female ; Humans ; Male ; Middle Aged ; Protamines ; blood ; Sensitivity and Specificity ; Thrombosis ; blood ; diagnosis

Background Corneal neovascularization ( CNV ) is a complication of many ocular surface diseases.It often worsen the pathological course.Effective therapy for CNV is still researching. Objective This study was to investigate the inhibitory effect of irradiation on CNV. Methods CNV models were established in 70 right eyes of 70 clean Wistar rats by corneal alkali burning.The models were randomized into β ray 10 Gy once irradiation group( 2 eyes),β ray 7 Gy multiple irradiation group( 17 eyes),β ray 10 Gy multiple irradiation group( 17 eyes),1％ cyclosporin A ( CsA ) eye drops group ( 17 eyes) and model group ( 17 eyes),and 6 matched normal rats were used as normal controls.All treatments started from the first day of the corneal alkali burning.CNV length and area were measured under the slit lamp every day.Corneal samples and homogenate were prepared 3,5,7 days after corneal alkali burning.The expressions of bcl-2,bax,vascular endothelial growth factor(VEGF) in rat corneas were detected by immunochemistry,VEGF proteins and VEGF mRNA were detected by Western blot and reverse transcription polymerase chain reaction (RT-PCR),respectively. Results Corneal ulceration was found in the βray 10 Gy once irradiation group and β ray 10 Gy multiple irradiation group.CNV length and area were much less in the β ray 7 Gy multiple irradiation group and 1％ CsA eye drops group compared with the model group on the seventh day after experiment( length:q=14.40,24.20,P＜0.01 ;area:q=17.80,14.00,P＜0.01 ).Immunochemistry revealed that compared with the model group,expressions of bcl-2 and VEGF proteins were weaker,but the expression of bax protein was stronger in the β ray 7 Gy multiple irradiation group and 1％ CsA eye drops group.RT-PCR showed that the expression of VEGF mRNA in cornea was lower in the β ray 10 Gy multiple irradiation group,β ray 7 Gy multiple irradiation group and 1％ CsA eye drops group in comparison with that in model group,and the results from Western blot showed the same pattern as RT-PCR. Conclusions Low dose irradiation of 90Sr-90Y ophthalmic applicator inhibits CNV formation after alkali burn.The study provide a new understanding of the irradiation for the treatment of CNV.