Since the first intein was found, more and more attenti on were paid on it. It not only enrichs the content of the process that the gene transfers its information but also can be used in protein purification. The rec ent advance in the sequence characteristic, transfer, evolution and the mechanis m of splicing of intein was summarized.

Objective To explore the value of intelligent optimum tube voltage technology at dual-source CT coronary angiography.Methods One hundred and fifty patients with normal body mass index (＜25 kg/m2) with clinically suspected coronary heart disease requiring coronary artery CTA examination were collected.According to the different scanning method,patients were randomly divided into 5 groups,30 cases of each group were tested by coronary artery CTA examination.The intelligent optimum tube vohage scanning technology was set to Semi,the tube voltage of A,B,C group were 120,100 and 80 kV (Ref 120 kV,400 mAs),respectively.D and E group were set to on,the reference voltage of group D was 120 kV,and reference current was 400 mAs.The reference voltage of group E was 100 kV,reference current was 400 mAs.Using the 15 section improved segment method for evaluation of coronary artery (lumen diameter＞ 2 mm and no severe calcification).They were assessed by subjective evaluation (image quality score) and objective evaluation (including vascular CT value,signal-to-noise ratio and contrast to noise ratio).Analyses of the differences between groups were compared with image quality,objective evaluation index and radiation dose by single factor variance.Results The image quality scores in patients scanned with five groups were (3.42±0.63),(3.41±0.54),(3.49±0.33),(3.45±0.43) and (3.48±0.81),there was no statistical difference between the five groups (F=0.634,P=0.105).Each image quality was good,all could clearly show thewhole range of vessels.The CT values of the coronary artery with above groups respectively were (486±82),(554±71),(742±90),(506±81),(561±81) HU.The image noises were (25±12),(32±12),(46±14),(28± 11),(34±12) HU (F=3.430 and 4.332,P＜0.05).And the SNR and CNR was no statistic difference between the five groups (P＞0.05).The effective radiation dose of five groups are (9.3± 1.3),(6.3± 1.4),(2.8±1.5),(5.7 ± 1.2),(3.9 ± 1.3) mSv,which the difference was statistically significant (F=2.332,P＜0.05).Conclusions Intelligent optimum tube voltage technology (Ref.100 kV,400 mAs) at dual-source CT coronary angiography may be feasible in patients with normal BMI.To ensure the quality of the image at the same time,it can effectively reduce the radiation dose.

A study on the microbial transformation of glycyrrhetinic acid (GA) was conducted by a fungus, Cunninghamella blakesleeana CGMCC 3.970 systematically. After incubation with the cell cultures of C. blakesleeana CGMCC 3.970 at 25 degrees C for 7 days on a rotary shaker operating at 135 r x min(-1), GA was converted into one major product and five minor products. The products were extracted and purified by solvent extraction, macroporous adsorbent resin, silica gel column chromatography, and semi-preparative RP-HPLC chromatography. Their structures were identified as 3-oxo-15α-hydroxy-18β-glycyrrhetinic acid(1), 3-oxo-15β-hydroxy-18β-glycyrrhetinic acid (2), 7β,15α-dihydroxy-18β-glycyrrhetinic acid (3), 3-oxo-7β, 15α-dihydroxy-18β-glycyrrhetinic acid (4), 7β-hydroxy-18β-glycyrrhetinic acid(5) and 15α-hydroxy-18β-glycyrrhetinic acid(6) by the analyses of MS, 1H-NMR and 13C-NMR spectroscopic data respectively. Among them, 2 was a new compound. These results suggest that C. blakesleeana CGMCC 3.970 has the capability of selective ketonization and hydroxylation for GA. [Key words] glycyrrhetinic acid; Cunninghamella blakesleeana CGMCC 3. 970; microbial transformation
Biotransformation ; Cunninghamella ; metabolism ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; metabolism ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo investigate the protective effect and mechanism of Shenqi Compound on diabetic angiopathy modeled rats.

METHODSTotally 18 SD rats were randomized into 3 groups, i.e., the normal control group, the diabetic mellitus (DM) group, and Shenqi Compound group, 6 in each group. The DM rat model was established by feeding high-fat diet (to induce hyperlipidemia) +intraperitoneal injection of small dose streptozotocin (STZ). Shenqi Compound was given to rats in the Shenqi Compound group at the daily dose of 2 g/kg. Equal volume of normal saline was given to rats in the model group and the normal control group by gastrogavage. All treatment was lasted for 12 weeks. Then 2-D and ultrasonic integrated backscatter technique were used to evaluate structural and functional changes of abdominal aorta in the progression of diabetic macroangiopathy. The fibrosis degree of the aorta vessel and myocardium capillaries were observed by using HE and Masson trichrome staining. The tension of the aortic vascular ring was determined. The transforming growth factor beta (TGF-beta) mRNA expression was detected by real time PCR (RT-PCR). The protein expression of TGF-beta, collagen I, collagen III, connective tissue growth factor (CTGF), and phosphorylation P38 MAPK were detected by Western blot.

RESULTSCompared with the normal control group, abdominal aortic systolic inner diameter, diastolic inner diameter, Peterson elastic modulus, stiffness index, and backscatter integral significantly increased; the rangeability of integral backscatter and the extension coefficient of cross section significantly decreased in the DM group (all P < 0.05). After 12 weeks aforesaid indices were obviously improved in the Shenqi Compound group (P < 0.05). Results of HE and Masson staining showed that the fibrosis degree of the aorta vessel and myocardium capillaries was obviously alleviated in rats of the Shenqi Compound group (P < 0.05). Results of the aortic vascular ring tension showed that acetylcholine induced vasodilatation and maximum diastolic percent were obviously elevated in the Shenqi Compound group (P < 0.05). Compared with the normal control group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all significantly increased in the DM group (P < 0.05). Compared with the DM group, the mRNA expression of TGF-beta, and the protein expression of TGF-beta, collagen I, and collagen III, and phosphorylation of P38 MAPK all decreased (P < 0.05).

CONCLUSIONSShenqi Compound could effectively improve the arterial function in diabetic marcoangiopathy and microvascular dysfunction. The mechanism might be due to the down-regulating the expression of TGF-beta, and further suppressing the phosphorylation of P38 MAPK, reducing the synthesis of collagen I and collagen III, therefore, ameliorating arterial and myocardial interstitial fibrosis.

Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Angiopathies ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta1 ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

The clinical tumor therapy was greatly challenged due to the complex characteristics of tumor microenvironment, however, which also provide arena for novel therapeutic strategies. In this study, poly(2-ethyl-2-oxazoline)-poly(lactic acid)-SS-poly(β-amino ester (PEOz-PLA-SS-PBAE) triblock copolymers with pH and GSH double response were synthesized, polymer micelles were prepared by thin film hydration method for loading of silybin to improve its antitumor activity. The critical micelle concentration was determined by pyrene fluorescence method as 1.8 μg·mL^-1. The particle size was 155.30 ± 1.80 nm as determined by dynamic light scattering, with polydispersity index of 0.168 ± 0.004. The drug loading and entrapment efficiency of the micelles were determined by HPLC as (5.48 ± 0.04)% and (68.52 ± 0.48)%, respectively. The in vitro drug release profiles showed that the micelles have low pH sensitivity and high GSH responsiveness, and exhibited sustained release profiles. The good biocompatibility of the material was proved by measuring the hemolysis rate and cytotoxicity of the blank micelle. The cytotoxicity and apoptosis rate of tumor cells showed that the drug loaded PEOz-PLA-SS-PBAE micelles had significant inhibitory effect and apoptosis-inducing effect on MDA-MB-231 cells. The results of wounding healing assay and Transwell invasion test showed that the drug loaded PEOz-PLA-SS-PBAE micelles could significantly inhibit the metastasis of MDA-MB-231 cells. The PEOz-PLA-SS-PBAE drug-loaded micelles prepared in this study have good inhibitory effect on tumor growth and anti-tumor metastasis in vitro, which lays the foundation for the further application of silybin.

Objective:To explore whether the deubiquitinating enzyme deubiquitin ligase 9X(USP9X)affects the proliferation of nasopharyngeal carcinoma(NPC)cells via regulating forkhead box Ml(FOXM1)mediated glycolysis. Methods:Western blotting and real-time fluorescence quantitative PCR,respectively,were used to detect the protein and mRNA expression levels of USP9X in NPC tissue and normal nasal mucosa tissue.NPC cells(CNE2 and HNE2)were infected with lentivirus carrying specific shRNA targeting USP9X gene(shUSP9X)or full-length USP9X gene(Flag-USP9X)for USP9X overexpression.CCK-8 assay was used to evaluate the effect of USP9X silencing or overexpression on the proliferation of NPC cells.Cellular energy metabolism assay and extracellular acidification rate(ECAR)assay were performed to investigate the impact of USP9X alteration on the glycolysis of NPC cells.The interaction between USP9X and FOXM1 was analyzed using ubibrowser_v3/database,co-immunoprecipitation,and ZDOCK software,and the changes in FOXM1 protein ubiquitination levels upon USP9X silencing was examined.Real-time fluorescence quantitative PCR and Western blotting,respectively,were used to detect the effect of USP9X alteration on the expression of FOXM1 mRNA and protein in NPC cells.Finally,FOXM1 expression was restored in USP9X silencing CNE2 cells by expression of recombinant plasmid Flag-FOXM1 carrying full-length FOXM1 gene through lentiviral infection.CCK-8 assay was used to evaluate the effect of FOXM1 upregulation on the proliferation of CNE2 cells.Cellular energy metabolism assay and ECAR assay were used to investigate the effect of FOXM1 upregulation on the glycolysis of CNE2 cells. Results:Compared with normal nasal mucosa tissue,the expression levels of USP9X mRNA and protein were significantly increased in NPC tissues(P＜0.05).After USP9X downregulation,the proliferation and glycolysis of CNE2 cells was significantly inhibited as indicated by the results of CCK-8 assay,cellular energy metabolism assay and ECAR assay(P＜0.05).In contrast,the proliferation and glycolysis of HNE2 cells was significantly enhanced after USP9X upregulation(P＜0.05).The interaction between USP9X and FOXM1 was confirmed by ubibrowser_v3/database,co-immunoprecipitation,and ZDOCK software analysis.Silencing USP9X could significantly increase FOXM1 ubiquitination in CNE2 cells.Overexpressing FOXM1 in low-USP9X CNE2 cells restored the proliferation and glycolysis activity of NPC cells(P＜0.05). Conclusion:USP9X can enhance NPC cell glycolysis by inhibiting FOXM1 ubiquitination and subsequent degradation,thereby promoting NCP cell proliferation.USP9X may be a potential novel therapeutic target for the treatment of NPC.

OBJECTIVETo study the types and emergence time of YMDD motif mutation in hepatitis B virus (HBV) polymerase gene during lamivudine treatment.

METHODSThe serum samples were collected from 33 patients with HBV DNA rebounding and 2 non-responders after at least one year lamivudine treatment. HBV polymerase gene was amplificated by PCR, then the products were detected by restriction fragment length polymorphism (RFLP) and by direct sequence analysis.

RESULTSThe variants with YMDD mutation were 14 out of the 35 patients. Mutation patterns detected in these patients included four YIDD, six YVDD, three YI/VDD and one YI/MDD. The mean emergence time of YMDD variants was 11.07+/-3.65 months after the treatment, and the earliest one and the latest one occurred 5 months and 17 months after the treatment respectively. The emergence times of YIDD, YVDD, YI/VDD were (10.00 +/- 1.41) months, (11.67 +/- 4.41) months and (13.33 +/- 3.31) months respectively, which had no statistical significance (F = 0.543, P < 0.05). Three patients treated with lamivudine 200 mg every day after the mutation were followed up for 6 months, whose HBV variants had not vanished.

CONCLUSIONSThere are many kinds of HBV variants after lamivudine treatment, including YIDD, YVDD, YI/VDD and YI/MDD. The emergence time of variants is quite variable between different types and the mean time is (11.07 +/- 3.65) months after treatment, and there is no relationship between the type of YMDD mutation and the time of lamivudine administration.

Adolescent ; Adult ; Aged ; Amino Acid Motifs ; genetics ; Child ; Cloning, Molecular ; DNA, Viral ; genetics ; Drug Resistance, Viral ; genetics ; Female ; Gene Products, pol ; genetics ; Hepatitis B virus ; drug effects ; enzymology ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Middle Aged ; Mutation ; RNA-Directed DNA Polymerase ; genetics ; Time Factors

Antiviral Agents ; therapeutic use ; DNA Mutational Analysis ; DNA, Viral ; blood ; Drug Resistance, Viral ; genetics ; Female ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Male ; Reverse Transcriptase Inhibitors ; therapeutic use

Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; etiology ; virology ; Humans ; Infant ; Rotavirus ; Rotavirus Infections ; epidemiology

OBJECTIVETo observe the clinical effect of hemiplegic muscle spasticity treated with acupuncture on different acupoints in combination with rehabilitation.

METHODSNinety cases of post-stroke muscle spasticity were randomized into a corresponding and central axis acupuncture group (group A), a conventional acupuncture group (group B) and a rehabilitation group (group C), 30 cases in each one. In group A, the acupoints on the Governor Vessel were mainly selected, as well as those on Hand-Shaoyang meridian and Foot-Taiyang meridian. In group B, the conventional needling and rehabilitation training were applied in combination, of which, the acupoints were mainly from Hand and Foot-Yangming meridians, associated with the acupoints of Shaoyang meridian. In group C, only rehabilitation training was applied. The assessments according to modified Ashworth scale and CSS score were performed before treatment, after 2 weeks and 4 weeks treatment respectively.

RESULTSThe level of modified Ashworth scale of the flexor of elbow and wrist was reduced apparently after treatment in group A as compared with that before treatment (P < 0.01). The level of modified Ashworth scale of the flexor of elbow was reduced apparently after treatment in group C as compared with that before treatment (P < 0.05). There was significant difference on the level change in modified Ashworth scale for the flexor of elbow between group A and group C after 4 weeks treatment (P < 0.05). CSS score decreased significantly in group A as compared with that before treatment (P < 0.01), the improvement after treatment in group A was apparent as compared with the other two groups (P < 0.01).

CONCLUSIONThe corresponding and central axis acupuncture can improve muscle tone of hemiplegic limb, which is superior to the effect achieved by the conventional acupuncture in combination with rehabilitation training and the simple rehabilitation training.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Aged, 80 and over ; Female ; Hemiplegia ; rehabilitation ; therapy ; Humans ; Male ; Middle Aged ; Muscle Spasticity ; rehabilitation ; therapy ; Treatment Outcome