Objective To investigate the clinical value of the probe melting curve analysis‐based PCR assay for the detection of three types of αT .Methods A total of 149 blood and prenatal archival DNA samples (6 of which were amniotic fluid samples)with three knownαT genes ,which included 63 carriers with Hb CS ,22 cases with Hb QS ,43 individuals with Hb WS and 1 double heter‐ozygote with Hb CS and Hb WS) as well as 20 samples with normalα‐globin gene sequence that had been confirmed by RBD com‐bined with DNA sequencing were selected to test the specificity of probe melting curve analysis by blind analysis .Results The probe melting curve analysis accurately detected 100 of the DNA samples previously characterized by S RBD combined with DNA sequencing .Conclusion Probe melting curve analysis‐based PCR assay for the detection ofαT is featured with rapidity and accuracy and can be applied to clinical and prenatal diagnosis .

Objective To investigate the reason of persisting positive rapid plasma reagin (RPR) in serofast syphilis patients, and to provide reference for clinical treatment and prognosis.Methods A total of 33 serofast patients and 23 healthy controls were enrolled in this study.The percentages and absolute counts of CD3+, CD4+, CD8+ T lymphocytes and natural killer (NK) cells were detected by flow cytometry.The comparison of two groups was analyzed by independent sample t test, and the correlation between change of lymphocyte subgroups and RPR titer in serofast syphilis patients was analyzed by bivariate linear correlation method.Results Compared with healthy controls, the percentages of CD3+, CD8+ T lymphocytes in serofast syphilis group were both increased significantly (75.75±5.76)% vs (68.37±5.80)%, (t=4.69, P<0.01);(27.34±7.02)% vs (24.33±1.95)%, (t=2.34, P=0.025), while both the percentage and absolute count of NK cells were significantly decreased (7.32±4.48)% vs (14.87±6.26)%, (t=5.269, P<0.01);(136.2±83.4)/μL vs (298.8±166.9)/μL, (t=4.311, P<0.01).RPR titer of the patients was negatively correlated with both percentage and absolute count of CD4+ T lymphocytes (r=-0.476 and-0.515, respectively, both P<0.01), and it was positively correlated of CD8+ lymphocytes (r=0.588 and 0.305, P<0.01 and P=0.804).Conclusion The imbalance of immune response of lymphocyte subsets observed in serofast syphilis may explain the RPR titers change.

A secretive expression vector of Pichia pastoris system which can be used for the direct cloning of PCR products was constructed,and was verified through the expression of recombinant cellobiohydrolase II in Pichia pastoris.A randomly selected fragment was amplified with properly designed primers by PCR.The XhoI and Eam1105Ⅰ restriction sites were included in the 5'end of the fragment,and the Eam1105Ⅰ and XbaI restriction sites were included in its 3'end.The PCR amplified product was inserted into the P.pastoris expression plasmid pPICZ?A through XhoI and XbaI restriction sites and the resultant plasmid was digested with Eam1105Ⅰ,and lastly the big fragment was recovered,generating the P.pastoris expressive Tvector pPICZ?T.Then the cellobiohydrolase II of T.reesei was successfully expressed in P.pastoris with this expressive Tvector.Such results indicated that the constructed expression Tvector was convenient for PCR product cloning,and was effective for heterologous protein expression in P.pastoris.On the other hand,the application of the expression Tvector avoided the introduction of additional amino acids at the Nterminus of the expressed protein,which generally occurred when normal expression vectors were used in secretive expression system.

As the main part of the teaching activities,students play an important role in the teaching reform.The students were trained through 3 pathways,“Extending teaching activities from the classroom to the outside”,“Development from basic to clinical knoledge” and “Culturing students' innovative consciousness”,so as to allow them to give full play in teaching reform,to enhance their ability of practice and learning by themselves,to culture their innovative consciousness and to develop students' leading functions in the anatomy teaching reform.

In order to select high quality and suitable Bupleuri Radix varieties in Qingchuan, and establish a new comprehensive method to evaluation the quality of Bupleuri Radix, 12 characters of 14 samples were evaluated by DTOPSIS and grey related degree. The results showed that varieties No. 8 and No. 10 had high quality. DTOPSIS and grey related degree gave the uniformity result, and the biggest difference of value of Ci in DTOPSIS method was 46. 33% , but the biggest difference of the weighting correlation number( r (i)) in grey related degree was only 13.10%. The DTOPSIS combined with grey related degree can evaluate the quality of Bupleuri Radix comprehensively and objectively.
Bupleurum ; chemistry ; China ; Drugs, Chinese Herbal ; chemistry ; supply & distribution ; Quality Control ; Statistics as Topic ; methods

Piroxicam has polymorphism. Different crystalline forms can exhibit different physicochemical properties and biological activities. Analysis of the intermolecular interactions is essential to reveal the formation mechanism and differences of polymorphs. In this paper, Hirshfeld surface analysis and semi-empirical methods were used to calculate and analyze the intermolecular interactions in seven polymorphic forms of piroxicam. The results show that the Hirshfeld surface analysis method can clearly and intuitively reveal the intermolecular interactions, among which H…H, O…H/H…O and N…H/H…N interactions account for 95% of the total energy. There are differences in the proportion and distribution of the forces of different crystal forms. The energy calculation shows that the lattice energy of the hydrate is significantly lower than that of the anhydrous forms, and in the specific energy distribution, the contribution of the dispersion force is the most prominent. Further interaction energy analysis was found that within the distance of 3.8 Å from the center of the piroxicam molecule, different crystalline forms of piroxicam molecule have different interaction energies with surrounding molecules.

The paper is aimed to study the difference in yield and quality at different harvest time and determine the optimum harvest of planting Gentiana in Ludian traditional harvest period. The authors analyzed the variation in fresh weight, dry weight, dry discount rate, length, diameter, volume and the content of gentiopicroside, loganin acid, alcohol-soluble extract and total ash and made a comprehensive appraisal of yield, appearance quality and intrinsic quality by gray relational distance ideal Comprehensive Evaluation method. The results showed that there is a big difference in yield and quality both 2-year-old and 3-year-old Gentiana harvested in traditional harvest period and the comprehensive evaluation more better when harvested more later. It can be seen, Gentiana harvested the later had a better yield and quality in Ludian traditional harvest period. The harvest of Gentiana can be appropriate delayed depending on the particular circumstances of production.
Agriculture ; methods ; China ; Gentiana ; anatomy & histology ; growth & development ; metabolism ; Iridoid Glucosides ; metabolism ; Organ Size ; Quality Control ; Time Factors

Objective To study antibiotic resistance phenotypes and genotypes of extended spectrum ?-lactamases (ESBLs) and AmpC ?-lactamase-producing Klebsiella oxytoca isolated from specimens of respiratory tract in children.Methods Bacterial isolates were identified by API or VITEK32. Agar dilution was used for antibiotic susceptibility test,and ESBLs and AmpC were detected by confirmatory test recommended by CLSI/NCCLS and by 3-aminophenylboronic acid (APB) disk potentiation test, respectively.Microarray was used to determine the genotypes of ESBLs and AmpC ?-lactamases.Genotypes of Klebsiella oxytoca were determined by enterobacterial repetitive intergenic consensus (ERIC)- PCR.Results ESBLs were positive in 129 out of 165 isolates (78.2%).Both ESBLs and AmpC ?- lactamases were positive in 16 out of 165 isolates (9.7%).AmpC ?-lactamase alone producer was not detected in term of phenotype and genotype.CTX-M was the most common type of ESBLs and DHA was the only type of AmpC ?-lactamase in these isolates.Most antibiotic resistant strains of Klebsiella oxytoca possessed the same genotype by ERIC-PCR.Although all strains were susceptible to carbpenem,Klebsiella oxytoca with ?-lactamases were more resistant to other antibiotic agents than those without ?- lactamases.Conclusions There is high prevalence of ESBLs production among Klebsiella oxytoca isolated from children in Urumqi.The main genotypes of ESBLs and AmpC ?-lactamases are CTX-M and DHA.

The Bcl-2 family of proteins play a central role in the control of apoptosis, a fundamental process for both human health and disease, by mitochondrial pathway. PUMA(p53 up-regulated modulator of apoptosis protein) is one of BH3-only members of Bcl-2 family , its function is to promote cell apoptosis. To obtain BH3 death domain peptide of PUMA and detect its biological activity, the synthesized double-stranded oligomeric nucleotide encoding PUMA-BH3 peptide was cloned into expression vector pTYB2,thus generating a construct of pTYB2-PUMA-BH3 which expressed PUMA-BH3-intein-chitin binding domain fusion protein. Then the recombinant plasmid was transformed into E.coli BL-21 (DE3) and fusion protein was expressed under induction by IPTG. The soluble PUMA-BH3 peptide was purified from chitin affinity chromatography by DTT reduction. Through measuring mitochondria viability(MTT),mitochondria permeability transition(MPT) and the translocation of cytochrome c(Cyt c ) assayed by western blotting, the biological pro-apoptotic activity of PUMA-BH3 peptide was studied. The PUMA-BH3 peptide has the effects on decreasing the mitochondria viability remarkably , inducing mitochondrial swelling and promoting Cyt c releasing from isolated mitochodria . Mitochondrial swelling and the release of Cyt c induced by PUMA-BH3 peptide concerned with the opening of MPT,which can be improved by cyclosporine A(CsA).These results indicated that recombinant PUMA-BH3 peptide might possess pro-apoptosis activity and paved a reasonable way for the study of new apoptosis regulators.

Objective To investigate the prevalence of mite sensitivity in patients with urticaria or other skin rashes, and to observe the clinical efficacy of a specific immunotherapy(SIT) by the Injectio dermatophagoidei farinae for the patients.Methods In 7-year period(1998-2005), skin prick test(SPT) with a dust mite(Df) allergen was carried out to detect the prevalence of mite sensitivity in OPD patients suffering from skin rashes.Among the patients sensitive to mite with SPT ≥++ response, 3 groups were established.In group A, routine SIT with Injectio dermatophagoidei farinae was conducted.In 9-week increasing dose phase, three stepwise increasing volumes(0.3ml, 0.6 ml and 1.0 ml) each case was injected subcutaneously with mite concentration of 1 ∶ 100 000(w/v) , 1 ∶ 10 000(w/v) or 1 ∶ 5 000(w/v) respectively once a week, followed by a maintenance dose phase for an injection with 1 ∶ 5 000(w/v) 1.0 ml/wk for 6 weeks.Group B received rush SIT with mite injections.A total of 15 injections in a course of therapy with same concentration and volume was given as those for the routine ones except shortened intervals, namely, 9 initial injections completed in 3 days by three injections of each concentration per day with two 30 min intervals, maintenancedoses were then provided in 6 days with 1 ∶ 5 000(w/v) 1.0 ml/d.Thereafter, both groups A and B were maintained for one year with a dose of 1 ∶ 5 000(w/v) 1.0 ml every 2 wk.Group C received antihistamine treatment as control, the patients received daily oral Ebastine 10 mg in the morning and Cetirizine dihydrochloride 10 mg in the evening for one week course and pro re nata later.Levels of serum tIgE and serum mite sIgE were detected by ELISA in 20 urticaria cases before and after one year mite SIT.Results Altogether, 2 685 cases with skin rashes were detected by Df allergen SPT.The prevalence of urticaria cases sensitive to mite was 70.3%(1 754/2 496), which was higher than that of eczema 63.5%(54/85) and anaphylactoid purpura 60.6%(63/104)(P