To achieve higher level expression of Interferon α2b (IFN-α2b) in methylotrophic yeast (Pichia pastoris), a cDNA fragment coding for the mature IFN-α2b was designed and synthesized based on the synonymous codon bias of P. pastoris and optimized G+C content. The synthetic IFN-α2b was inserted into the secreted expression vector pPICZαA, and then integrated into P. pastoris GS115 genome by electroporation. Multi-copy integrants in the Mut+ recombinant P. pastoris strain were screened by high concentrations of Zeocin. 120 hours culturing allowed expression of the IFN-α2b transformant up to 810 mg/L as detected by SDS-PAGE and quantitative methods. In addition, Western blot analysis showed that the recombinant proteins had immunogenicity. The significant antiviral activity of the recombinant IFN-α2b protein was verified by WISH/ VSV system, which was 3.3×105 IU/mL.

Objective To investigate the value of functional MRI in the diagnosis and differential diagnosis of breast diseases.Methods Sixty-five patients with 68 lesions were enrolled in this study. Conventional T_1 WI and T_2 WI scan,dynamic contrast enhanced MRI,diffusion weighted imaging and ~1H single voxel MR spectroscopy were performed consequently.All lesions were verified by pathology,including 4 cases of breast adenosis,22 fibroadenomas,2 chronic inflammations,3 cysts,33 infitrating ductal carcinomas,1 intraductal carcinoma and 3 cystosarcoma phyllodes tumors.Morphological features,maximum enhancement ratio,time-intensity curve,apparent diffusion coefficient and Choline peak were analyzed. Results The detection rates of T_1 WI and T_2 WI were 14.7%(n=10)and 51.5%(n=35).The sensitivity,specificity,accuracy of dynamic contrast.enhanced MRI for the malignant tumor were 94.6%, 71.4% and 76.5% respectively.Retrospective study showed that diffusion weighted imaging,with the b value from 800 s/mm~2 to 1000 s/mm~2,could be used to differentiate various types of breast lesions.~1H signal voxel spectroscopy had a sensitivity of 51.4%,specificity of 82.6%,and accuracy of 67.6% for the malignent.The sensitivity,specificity and accuracy could reach 97.3%,90.0% and 92.6% respectively by combining conventional scan,dynamic contrast enhanced MRI and MR spectroscopy.Conclusion Functional MRI,with high sensitivity,specificity and accuracy,can be used widely in the diagnosis of malignant breast lesions.

Isolated alkaloids from Coptis chinensis Franch. The compounds were identified as berberine, columbamine, groenlandicine, jatrorrhizine, magnoflorine, corydaldine and ferulic acid methylester. Then measured their bitter degree based on the electronic tongue and evaluated the antibacterial. The results based on the Electronic Tongue showed that berberine, columbamine, groenlandicine and jatrorrhizine have higher bitter degree than magnoflorine and corydaldine. And they also appeared better antibacterial activity on E. coli and S. aureus. The correlation coefficients between bitter degree and the two bacteria antibacterial activity were 0.983 and 0.911. So there was close relationship between the bitter degree and antibacterial activity of bitter components. Thus, it is confirmed further that bitter components are the material foundation of medicinal effectiveness of bitter herbs.
Aporphines ; analysis ; Berberine ; analogs & derivatives ; analysis ; Berberine Alkaloids ; analysis ; Biomedical Research ; instrumentation ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; analysis ; chemistry ; pharmacology ; Electronics ; instrumentation ; methods ; Escherichia coli ; drug effects ; growth & development ; Microbial Sensitivity Tests ; Reproducibility of Results ; Staphylococcus aureus ; drug effects ; growth & development ; Taste

Child, Preschool ; Fatal Outcome ; Female ; Humans ; Respiratory Tract Infections ; complications ; Shock, Septic ; etiology ; Streptococcal Infections ; complications

OBJECTIVETo observe the effect of Ningdong Granule (NG) on serum levels of interleukin-12 (IL-12) and tumor necrosis factor-alpha (TNF-alpha) of children patients with Tourette's syndrome (TS).

METHODSTotally 90 TS children patients were randomly assigned to the NG group, the NG + Tiapride group (abbreviated as the combined treatment group), and the Tiapride group, 30 in each group. Besides,another 30 healthy children were recruited as the healthy control group. Patients in the NG group were treated with NG (consisting of all gastrodia rhizome, Codonopsis pilosula, Ophiopogon japonicus, white peony root, Rhinocerotidae, oyster, earthworm, licorice root, etc.), one dose daily, administered by dissolving it in boiled water, taken in two portions in the morning and in the evening respectively. Patients in the Tiapride group took Tiapride Tablet, 50 -100 mg each time, twice daily. The dosage was adjusted according to individual difference and changes of pathogenic conditions. The maximal dosage was 300 mg per day. Those in the combined treatment group were treated with equal dose of NG and Tiapride Tablet in combination. The treatment course was 3 months for all. Changes of pathogenic condition before and after treatment were assessed by Yale global tic severity scale (YGTSS). Serum levels of IL-12 and TNF-alpha were detected by enzyme-labeled immunosorbent assay (ELISA) before and after treatment.

RESULTS(1) The total effective rate of the NG group, the combined treatment group, and the Tiapride group was 79.3%, 83.3%, and 67.9%, respectively. It was the lowest in the Tiapride group (P < 0.05). It was significantly higher in the combined treatment group than in the NG group (P < 0.05). (2) The post-treatment YGTSS score was obviously lower in each group after treatment than before treatment (P < 0.05). The posttreatment YGTSS score was obviously lower in the NG group and the combined treatment group than in the Tiapride group (P < 0.05), but with no statistical difference between the fromer two groups (P > 0.05).(3) Compared with the healthy control group before treatment, serum levels of IL-12 and TNF-alpha (pg/mL) were 124.95 +/- 22.78 and 209.52 +/- 21.69 in the NG group, 126.14 +/- 25.65 and 208.97 +/- 22.46 in the combined treatment group, 123.00 +/- 24.26 and 205.10 +/- 26.16 in the Tiapride group, being higher than those in the healthy control group (64.56 +/- 27.59 and 78.13 +/- 33.42; P < 0.05). After treatment, serum levels of of IL-12 and TNF-alpha were 104.67 +/- 16.84 and 183.01 +/- 24.95 in the NG group, 109.04 +/- 16.81 and 179.87 +/- 23.45 in the combined treatment group, significantly lower than before treatment (P < 0.05). But there was no statistical difference in serum levels of IL-12 or TNF-alpha in the Tiapride group between before treatment (123.00 +/- 24.26 and 205.10 +/- 26.16) and after treatment (117.75 +/- 16.79 and 199.76 +/- 33.21; P > 0.05).

CONCLUSIONNG could modulate abnormal serum levels of IL-12 and TNF-alpha in TS children patients, which might be one of its pharmacodynamic mechanisms for treating TS.

Adolescent ; Child ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-12 ; blood ; Male ; Phytotherapy ; Tourette Syndrome ; blood ; drug therapy ; Tumor Necrosis Factor-alpha ; blood

Background The tissue-engineered cornea is becoming the hot spot in the ophthalmologic field,while the research of corneal substitute is in the ascendant,because it is more similar to the corneal morpha and easy to survive in vivo.Objective This study was to investigate the biocompatibility of recombinant human type-Ⅲ collagen/poly9 ( 3-( methacryloylamino ) propyl dimethyl ( 3-sulfopropyl ) ammonium hydroxide ) ( PMPDSAH ) interpenetrating polymer network (IPN) (RHC-Ⅲ/PMPDSAH IPN) hydrogel as a tissue-engineered cornea in rabbit eye and its feasibility as the corneal substitute.Methods One hundred and eight rabbits were randomly divided into experimental group( 90 rabbits) and normal control group ( 3 rabbits),and 15 rabbits ( 30 eyes ) used as the donor corneas.RHC-Ⅲ/PMPDSAH IPN,NGF PMPDSAH IPN and corneal grafts were lamellarly transplanted into the right eyes in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group respectively.The corneal transparency and neovascularization were examined and scored under the slim lamp and compared among three groups using Kraskal-Wallis H test.The corneal epithelization time was observed and compared among these three groups using one way analysis of variance and LSD-t test.The histological examination of corneas was performed at the 3rd day,1st and 2nd week,1 st,3rd and 6th month after the surgery.The immunohistochemistry was used to detect the expression of K3 in cornea at the 6th month.Results The grafts were well attached in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group,and no rejection reaction was found throughout 6-month following up.Compared with normal control group,no significant differences were found in the scores of corneal opacification and neovescularization in these three groups (x2 =4.34,P =0.23 ;x2 =2.60,P =0.46 ) at the 6th month.NGF PMPDSAH IPN group achieved reepithelialization in (4.97±0.63) days and was obviously shorted than that in RHC-Ⅲ/PMPDSAH IPN group and allograft group ( t =11.97,P =0.00; t =5.80,P =0.00).The re-epithelialization time in RHC-Ⅲ/PMPDSAH IPN was (6.86±0.71) days,and that of allograft group was (5.87±0.43 ) days,showing a significant difference ( t =6.32,P =0.00).Hematoxylin-eosin staining results demonstrated that implanted materials integrated into the host corneal tissue well and support corneal epithelialization.Part of the material degraded at the 2nd week and degraded completely 1 month later.Regular alignment and distribution of collagen fibers were seen in the regenerated cornea and were similar to those of the normal stroma in 6 months.Immunohistochemistry showed the positive expression of keratin-3 in corneal epithelial cells.Conclusions RHC-Ⅲ/PMPDSAH IPN has a good biocompatibility without toxicity to corneal tissue.Furthermore,NGF can promote the corneal wound-healing and re-epithelialization.The material can be used as safe and reliable corneal substitute after improving the mechanical strength.

Objective To investigate the diagnosis value of immunophenotype and karyotypes in newly diagnosed chronic lymphocytic leukemia (CLL).Methods To retrospect the flow cytometry (FCM) immunophenotype and karyotypes characteristics in newly diagnosed 70 CLL cases.Results In all cases,the positive rates of CD19,CD20,CD5,CD23,CD22 were 100 ％,88.5 ％ (54/61),77.1％ (54/70),67.6 ％ (23/34)and 51.9 ％ (27/52),respectively.And 6 were misdiagnosed,2 was CD+5CD+19(+),but CD20,CD22 were strongly positive,final diagnosed as mantle cell lymphoma (MCL) by FISH t(11;14) examination and CyclinDl; CD+5CD+19(-) CLL were 16 cases (22.9 ％),but 4 were misdiagnosed,the misdiagnosis rate was 25 ％,significantly higher than that of CD+5CD+19(-) CLL (P =0.030).59 cases were examined by conventional cytogcnetic (CC),and 13 cases were with abnormal karyotypes,positive rate was 22.0 ％,with complex karyotypes in 5 cases (8.5 ％); 10 cases combined with FISH abnormalities karyotype examination rate was 60 ％.Conclusion Typical CLL immunophenotypic characteristics were CD5,CD19,CD23 co-expression,and CD－5 CLL with higher misdiagnosis rate,combined with CD20 (,) CD22 expression and karyotype analysis helps to CLL and other B lymphoid proliferative diseases (B-LPD) identification.Conventional cytogenetic detection combined with FISH scan can improve the recognition ability of abnormal chromosome.

Objective To compare the efficacy of 18F-FDG PET/CT, 99Tcm-MDP bone scintigraphy (BS), and combination of the two techniques (PET/CT + BS) for detecting bone metastasis by ROC curve analysis. Methods All 296 patients with various cancers, who underwent both 99Tcm-MDP BS and 18F-FDG PET/CT within two months, were retrospectively analyzed. These images were interpreted according to 5-point scale (0: definitely negative, 1: probably negative, 2: equivocal, 3: probably positive, 4:definitely positive for bone metastasis), and the scale of PET/CT + BS was the sum of PET/CT and BS. In light of the confirmed diagnosis derived from pathology or follow-up, ROC curve analysis was performed.The area under the ROC curve (AUC) was compared by z-test. Results Of 296 cases, 61 (20.6%) were confirmed as bone metastases and 235 (79.4%) were negative. The AUC were 0. 919 (95% confidence interval (95% CI) :0. 867 - 0. 971) for BS, 0. 949 (95% CI: 0. 906 - 0. 991) for PET/CT, and 0. 994 (95% CI: 0.988-0.999) for PET/CT + BS, rctrospectively. The AUC of PET/CT + BS was statistically significantly larger than that of BS (z=2. 866, P=0.004) or PET/CT (z =2.027, P=0.043), while the AUC of PET/CT was larger than that of BS, but no statistically significance (z = 0. 881, P = 0. 378) was showed. The optimal sensitivity, specificity, accuracy, positive predictive value (PPV), negative predictive value(NPV) were 90. 2% (55/61), 85. 1% (200/235), 86. 1% (255/296), 61. 1% (55/90), 97. 1%(200/206) for BS, 88.5% (54/61), 97.0% (228/235), 95.3% (282/296), 88.5% (54/61), 97.0% for PET/CT, and 98.4% (60/61), 95.7% (225/235), 96.3% (285/296), 85.7% (60/70) for PET/CT + BS,respectively. The specificity (χ2 = 19.862, P<0. 001), accuracy (χ2 = 23. 361, P<0.001) and PPV (χ2 =11. 791, P =0.001) of PET/CT + BS were significantly higher than those of BS, the sensitivity of PET/CT +BS was significantly higher than that of PET/CT (χ2 =4.167, P=0.031). Compared with BS, PET/CT had a higher specificity (χ2 = 19.600, P<0. 001), accuracy (χ2 = 13. 755, P <0. 001), PPV (χ2 = 13. 608, P <0. 001), but their sensitivity showed no statistically significant difference (χ2 = 0, P = 1. 000). Conclusions The efficacy of 18F-FDG PET/CT for detecting malignant bone metastasis was superior to that of 99Tcm-MDP BS alone. The detection ability can be obviously improved by combination of the two techniques.

Rats were continuously given different doses of water extract of Polygonum multiflorum (1, 10 g x kg(-1)) for 7 days to prepare liver microsomes. Cocktail in vitro incubation approach and Real-time quantitative PCR technology were used to observe the effect of water extract of P. multiflorum on CYP450 enzymatic activities and mRNA expressions in rat liver. Compared with the blank control group, both 1, 10 g x kg(-1) water extract of P. multiflorum treated groups showed significant inhibitions in CYP2E1 enzymatic activities and mRNA expressions (enzymatic activities of CYP2E1, P < 0.01; mRNA expression of CYP2E1, P < 0.05 in 1 g x kg(-1) group, P < 0.01 in 10 g x kg(-1) group). They revealed a significant increase in the enzymatic activity of CYP3A1 (P < 0.01), but without significant change in mRNA expressions. The 10 g x kg(-1) group showed a significant inhibition in CYP1A2 enzymatic activities and mRNA expressions in rat livers (P < 0.01).
Animals ; Cytochrome P-450 CYP1A2 ; genetics ; metabolism ; Cytochrome P-450 CYP2E1 ; genetics ; metabolism ; Cytochrome P-450 Enzyme Inhibitors ; administration & dosage ; isolation & purification ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; Liver ; drug effects ; enzymology ; Male ; Microsomes, Liver ; drug effects ; enzymology ; Polygonum ; chemistry ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo discuss the perioperative management of hemophiliacs A with orthopaedic complications.

METHODSTo regulate the injection of factor VIII concentrate in peroperative period by testing the level of factor VIII:C in 27 cases. The lever of factor VIII:C was improved to 30% - 50% at the day before the operation. To the severe patients, it was maintained at 58.5% - 89.3% during the operation and at 47.0% - 78.4% in postoperation. While to the gentle, it was maintained at 38.5% - 52.5% during the operation and at 29.2% - 52.3% in postoperation. The individualized surgical procedures were carried out, such as arthrocentesis, open knee synovectomy or arthroscopic synovectomy, evacuation or curettage of haematoma, debridements, internal fixation.

RESULTSAll patients were cured in 14 - 105 days by regulating the injection of factor VIII during the peroperative period. The usage of factor VIII was 1 200 - 70 250 IU.

CONCLUSIONWhile the hemophiliacs have orthopaedic complications, it is necessary to promise the factor VIII:C being maintained at the lever of hemostasis during the peroperative period. The hemophiliacs are endurable to the operation.

Adolescent ; Adult ; Factor VIII ; administration & dosage ; Fractures, Bone ; complications ; surgery ; Hemarthrosis ; etiology ; surgery ; Hematoma ; etiology ; surgery ; Hemophilia A ; blood ; complications ; drug therapy ; Humans ; Male ; Perioperative Care ; Postoperative Complications ; blood