AIM: To establish the method for quality control of Guipi Pills(concentrated pill) (Radix Codonopsis,Rhizoma Atractylodis macrocephalae,Radix Astragali,Radix et Rhizoma Glycyrrhizae,etc).and a quantitative method of determining astragaloside IV. METHODS: Radix Codonopsis,Radix Angelicae Sinensis,Radix et Rhizoma Glycyrrhizae and Radix Aucklandiae in the pill were identified by TLC.Astragaloside IV in the pill was determined by HPLC-ELSD. RESULTS: For astragaloside IV,the linear range was within 25.4-508.3 ?g and the average recovery was 99.1%,r=0.999 7 with RSD of 0.8%. CONCLUSION: The method is simple,reliable,accurate,practical.It can be used in the quality control of Gui Pipills(concentrated pills).

Objective To explore the change of copeptin in plasma and its significance in patients with intracerebral hemorrhage combined with stress ulcer.Methods Eighty patients with intracerebral hemorrhage were collected.Forty-nine patients of pure intracerebral hemorrhage and 31 patients of intracerebral hemorrhage combined with stress ulcer were included.Thirty healthy people were taken as controls.The level of copeptin in plasma was measured and compared in all subjects.Results The level of copeptin in plasma in patients with pure intracerebral hemorrhage and intracerebral hemorrhage combined with stress ulcer was significantly higher than that in controls:(303.684 ± 68.691),(527.034 ± 74.111) ng/L vs.(121.460 ± 53.364) ng/L,and the level of copeptin in plasma in patients with intracerebral hemorrhage combined with stress ulcer was significantly higher than that in patients with pure intracerebral hemorrhage.The differences were statistically significant (P ＜ 0.05).Conclusion The level of copeptin in plasma in patients with pure intracerebral hemorrhage increases significantly,and it is much higher in patients with intracerebral hemorrhage combined with stress ulcer.

Objective To observe the curative effect of methylprednisolone combined with entecavir in treatment of hepatitis B virus (HBV) related early stage liver failure.Methods One hundred and twenty-six patients with HBV related early stage liver failure were divided into treatment group (68 cases) and control group (58 cases) by random digits table method.The patients in 2 groups were given conventional hepatinica treatment and entecavir antiviral treatment,but the patients in treatment group were added methylprednisolone and pantoprazole.The alanine aminotransferase (ALT),total bilirubin (TBil),albumin,prothrombin time (PT),HBV-DNA,tumor necrosis factor (TNF)-α,interleukin (IL)-6 levels were compared between 2 groups,and the adverse reaction of methylprednisolone was observed.Results The ALT,TBil,PT and albumin levels after the first,second,fourth,sixth and eighth week of treatment in treatment group were significantly better than those in control group,and there were statistical differences (P ＜ 0.05).There was no statistical difference in HBV-DNA between 2 groups (P ＞ 0.05).The TNF-α and IL-6 levels after the first and second week of treatment in treatment group were (4.13 ± 1.25) and (1.98 ± 0.67) p g/L,(3.21 ± 0.75)and (1.23 ± 0.29) μ g/L,and in control groups were (5.89 ± 1.78) and (3.67 ± 0.87)μ g/L,(4.12 ± 0.88) and (2.68 ± 0.81) μ g/L.The TNF-α and IL-6 levels in treatment group were significantly lower than those in control group,and there were statistical differences (P ＜ 0.05).The effective rate in treatment group (79.41％,54168) was significantly higher than that in control group (51.72％,30/58),the fatality rate in treatment group (2.94％,2/68) was significantly lower than that in control group (24.14％,14/58),and there were statistical differences (P ＜ 0.05).The adverse reaction of methylprednisolone in treatment group was not found.Conclusion The methylprednisolone combined with entecavir can improve liver function and survival rate in patients with HBV related early stage liver failure,and adverse reaction of methylprednisolone is rare.

Objective To evaluate the outcomes of small (5 ～ 10mm),arterially enhancing nodules (SAENs)shown exclusively at the hepatic arterial phase of CT in a hepatocellular carcinoma(HCC) surveillance population and to determine risk factors for developing HCC.Methods The study population included 112 patients (maleifemale =100 ∶ 12 ; aged 36 ～ 92 years) with 1 7 5 SAENs who were at risk of HCC.We evaluated serial changes during follow-up(1.4 ～41.8 months,mean 35.7 months) and analysed the initial CT findings of SAENs and the accompanying lesions to elucidate the risk factors for HCC development.Results Of 175 SAENs,101 (57.7％) disappeared and 34(19.4％) persisted.Forty SAENs(22.9％) became HCC in 33 patients(29.5％).Presence of HCC treatment history(P =0.005,risk ratio =7.429),a larger size of SAEN(P =0.003,risk ratio =1.630),presence of coexistent HCC(P =0.021,risk ratio =3.777) and absence of coexistent typical arterioportal shunts (P =0.003,risk ratio =4.459) turned out to be independently significant risk factors for future development of HCC.Conclusion SAENs are frequently seen in an HCC surveillance population and have a 22.9％ probability of becoming HCC on a per-lesion basis.Risk increased particularly when the lesion is associated with a previous or concurrent HCC,a large size or found without a coexistent typical arterioportal shunt.

Objective To study the influence of Naikan therapy on the family function in patients with anxiety disorder of cerebral infarction.Methods A total of 192 patients with anxiety disorder of cerebral infarction were divided into control group and study group according to the set sequence.The study group was treated with Naikan therapy and the control group received routine therapy.All patients were assessed by family assessment device (FAD),Hamilton depression scale (HAMD),and Hamilton anxiety scale (HAMA).Results After 6-week treatment,the HAMD,HAMA,the affective reaction and behavior control scores in the study group were significantly lower than those before the treatment (t =2.24,2.43,2.37,2.35,P ＜ 0.05).The affective reaction and behavior control in the study group were obviously lower than that of the control group (t =2.51,2.41,P ＜ 0.05).Conclusion Naikan therapy can improve family function in patients with anxiety disorder of cerebral infarction.

Objective To study the influence of Naikan therapy on depression in patients with Schizophrenia.Methods 256 patients with Schizophrenia were divided into study group and control group according to the set sequence.The study group was simulated by Naikan therapy and the control group was given routine cure.All patients were assessed by Positive and negative symptom scale (PANSS),Simple coping style questionnaire (SCSQ),Defense way questionnaire (DSQ),Hamilton depression scale (HAMD),and Hamilton anxiety scale (HAMA).Restlts In the 6th week,the differences in HAMD score,Cover factor,negative coping were statistically significant between the two groups (t=2.37,2.33,2.36,P＜0.05).The differencs in HAMA score,Cover factor,negative coping were statistically significant between the 0th and the 6 th week in the study groups (t =2.31,2.42,2.47,P＜ 0.05).The difference of PANSS score were statistically significant between the 0th and 6th week of the two groups.Concltsion The Naikan therapy can improve the mental symptoms and depressive symptoms in patients with Schizophrenia.

AIM:To investigate the relationship of clinical efficacy,blood trough concentration and adverse reavtion,which in the treatment of methicillin-resistant Staphylococcus aureus infection with Vancomycin.METHODS:The clinical data of 82 hospital patients who were infected by methicillin-resistant Staphylococcus aureus and treated with Vancomycin from January 2008 to October 2009 were retrospectively analyzed the clinic efficacy,bacteriological treatment,adverse reaction were observed,and the blook trough concentration was detected.RESULTS:The treatment effective rate of Vancomycin-blood trough concentration which was less than 5 ?g/mL,in the range of 5-10 ?g/mL,and greater than 10 ?g/mL were 68.75%,72.73% and 81.82%.The bacterial clearance rates were 62.50%,68.18% and 72.73%.One case of blood trough concentration of 14.6 ?g/mL in patients with renal toxicity.CONCLUSION:Increasing the blood trough concentration of Vancomycin can improve the treatment of methicillin-resistant Staphylococcus aureus infection in the efficacy and bacterial eradication rates,but the probability of renal toxicity adverse reactions will also increase accordingly.

The anti-hepatitis B virus (HBV) effects and its mechanisms of the ethanol extracts of Hypericum perforatum L. (EHP) in vitro were explored. HepG2 2.2.15 cells, a stable HBV-producing cell line, were cultured as the model system to observe the anti-HBV effect. The viral antigens of cellular secretion, HBsAg and HBeAg, were determined by enzyme linked immunosorbent assay (ELISA). The quantity of HBV-DNA released in the supernatant was assayed by real-time PCR. In order to understand the mechanisms of the suppression of HBV replication, all HBV promoters (Cp, Xp, S1p, S2p and Fp) with luciferase reporter gene were transfected into HepG2 cells respectively. Then the activities of viral promoters were examined by luciferase reporter assay. It was found EHP effectively suppressed the secretion of HBsAg and HBeAg from HepG2 2.2.15 cells in a dose-dependent manner, as well as the extracellular HBV DNA. And EHP could selectively inhibit the activity of HBV promoter Fp. Our data suggest that EHP exerts anti-HBV effects via inhibition of HBV transcription, which helps to elucidate the mechanism underlying the potential therapeutic value of EHP.

ObjectiveTo understand the characteristics of mutations in BCR-ABL1 kinase domain mutation,these chronic myeloid leukemia (CML) and Ph positive acute lymphoblastic leukemia (ALL)patients who got imatinib treatment had poor effect.MethodsTotally 177 CML patients and 33 Ph( + )ALL patients were selected at Beijing Dao-Pei Hospital from Sep.2007 to Dec.2010.All of them were Chinese patients.Totally 243 bone marrow or peripheral blood specimens were collected from the patients,who had early effect,then resistance emergenced,or for more than 3 months of poor efficacy.Extracted total RNA from the specimens' nuclear cells,reversed transcription to cDNA.Amplified the whole span of BCRABL1 fusion kinase gene by nest PCR (from 242 to 493 amino acid coding sequence),used the type AB3130XL gene sequencing instrument determinate the gene sequence of ABL1 kinase region and then used the Variant Reporter V1.0 software to analyze the results of gene mutations.ResultsThirty-two kinds of different mutations were detected of ABL1 gene mutations,accounting for 34.2％ (83/243 cases).Among them,the T315I was 12％ (10/83),mutation rate was the highest,followed by Y253H was 11％ (9/83),G250E was 7％ (6/83),E255K was 7％ (6/83),M351T was 6％ (5/83),E459K was 5％ (4/83) ;Q252H,D276G,F317L,E355G,F359V,H396R were all 4％ (3/83).Three cases of insertion mutations were found,including 2 cases of 357-358insk,1 case of V304RfsX17.Seven patients had found existence two or more point mutations.The multiple drug resistance mutations might exist in the same leukemia clone.The same individual was not only contain common resistance mutations,but also rare point mutations,insertion mutations.The mutations might be lead to loss of kinase activity.ConclusionsUnder the imatinib drugs pressure,the ABL1 gene mutation in leukemia cells appears randomly,and results in different resistant clones.Different resistant clones can coexist in the same patients in vivo; resistant clones not only contain point mutations,but also contain inserted deletion mutations.

To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
Animals ; Animals, Genetically Modified ; genetics ; Female ; Fibroblasts ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Goats ; genetics ; Humans ; Lactoferrin ; genetics ; Lactoglobulins ; genetics ; Milk ; chemistry ; Nuclear Transfer Techniques ; Plasmids ; Pregnancy ; Transfection