To improve bond selectivity of recombinant β-glucuronidase in Escherichia coli (PGUS-E), based on the PGUS-E structure guidance, three key points R329, T369 and N467 were identified to be responsible for the bond selectivity of PGUS-E, and further saturation mutagenesis was conducted. Two positive mutants R329K and T369V were obtained by a combined selection technique of thin-layer chromatography and high performance liquid chromatography. Compared to PGUS-E, the bond selectivity of mutants R329K and T369V increased by 26.9% and 34.3%, respectively; whereas the biochemical properties such as pH and temperature profile were unchanged. Nevertheless, the activity was decreased compared to PGUS-E. These results further confirmed that sites R329 and T369 played important roles for the bond selectivity and activity. In summary, this study significantly increased the bond selectivity of PGUS-E by structure guided saturation mutagenesis, providing experimental support for elucidating the relationship between the structure and function of PGUS-E.
Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Escherichia coli ; metabolism ; Glucuronidase ; chemistry ; Industrial Microbiology ; Mutagenesis ; Recombinant Proteins ; chemistry ; Temperature

Cell Line, Tumor ; Enzyme Inhibitors ; pharmacology ; Heterocyclic Compounds, 1-Ring ; pharmacology ; Humans ; Proteasome Endopeptidase Complex ; drug effects

Objective To investigate the result and side effect of late course accelerated three-di-mensional conformal radiotherapy (3DCRT) for esophageal carcinoma. Methods From July 2003 to March 2006, 55 patients with esophageal carcinoma receiving 3DCRT were randomly divided into late course accel-erated radiation group (group A, 27 patients) and conventional fractionation group (group B, 28 patients). The prescribed dose in group B was 64 -66 Gy, 2 Gy per fraction, 1 fraction per day, 5 fractions per week for about 6.5 weeks. Patients in group A received conventional fractionation irradiation for the first 4 weeks. Then the dose was increased to 3 Gy per fraction to a total dose of 67 -70 Gy. The treatment course in group A was about 6 weeks. The treatment response, acute site effects, 1-, 3-and 5-year local control rates and o-verall survival rates of the two groups were observed. Results In group A, 23 patients (85%) achievedcomplete response (CR) and 4(15%) achieved partial response (PR). While in group B, 16 patients (57%) achieved CR and 12(43%) achieved PR. The CR rate was significant higher in group A (χ~2 = 5.24,P=0.022). The 1-, 3-, 5-year local control rates were 85%, 54%, 54% in group A, and 70%, 56%, 33 % in group B (χ~2 = 0.68, P = 0.409), respectively. The 1 -,3-,5-year overall survival rates of the two groups were 81%, 37%, 29% and 61%, 39%, 23% (χ~2 = 0.06, P = O. 804), respectively. Both lo-cal control and overall survival were similar between the two groups. The incidences of acute radiation esoph-agitis in the two groups were similar (85% vs. 89% ;χ~2 =0. 00,P=0. 959), and the incidence of radiation pneumonitis was slightly higher in group A than in group B (67% vs 43% ;χ~2 =3.14,P =0.076). By the last follow up, 19 patients in group A and 21 in group B died. Among them, 10 in group A and 15 in group B died of local failure, while 7 in group A and 5 in group B died of metastasis. Conclusions When com-pared with conventional fractionation 3DCRT, late course accelerated 3DCRT for esophageal carcinoma can achieve better results in clinical response, though not in long-term local control or survival. The incidence of acute radiation esophagitis and pneumonitis is clinically acceptable.

Objective To observe the incidence of RP in NSCLC and esophageal carcinoma treated with 3DCRT and investigate the relationship between acute RP and lung function and dosimetric parameters.Methods From October 2006 to August 2008, 3DCRT plus concurrent chemotherapy of NP or LFP were applied to 64 patients with locally advanced NSCLC or esophageal carcinoma. twenty-three patients suffered form NSCLC and 41 patients from esophageal carcinoma, the prescription doses were 60 Gy/30fx and 58 -64 Gy/29 -32fx, respectively. Results For patients with esophageal carcinoma, 34% developed RP(9 grade 1,3 grade 2 and 2 grade 3). For patients with NSCLC, 96% developed RP(9 grade 1, 8 grade 2 and 5 grade 3). There was significant difference between the two groups(t =5. 55,P=0. 000). The FEV1.0/FVC and DLCO of patients with NSCLC were significantly lower than those of esophageal carcinoma, the ratio were 75.6%:82.7%(t=2.75,P=0.008)and 71.7%:81.0%(t=2.50, P=0.015),respectively. For patients whose FEV1.0, FEV1.0/FVC%, DLCO ＜80% and ≥80% before irradiation,the incidence of ≥2grade ARP were 35% vs 25% ,31% vs 26% and 35% vs 19%, respectively(x2 = 1.81,0.15,2. 13,P =0.179,0.697,0.144). While for patients whose FEV1.0 ＜ 70% and ≥70%, the incidence of severe ARP were 67% and 22% ,respectively(x2 =5.64, P =0.018). Spearman correlated analysis indicated that all the dosimetric parameters had relation with ≥ 2 grade ARP . The V20 of lung and MLD were found independently associated with RP according to multivariate analysis(x2 = 4.61,6.97, P = 0.032,0.008).Conclusions Parameters of basic lung function can predict the incidence of ≥2 grade RP to some extent,especially when the value of FEV1.0, FEV1.0/FVC%, and DLCO was lower. However, the V20 of lung and MLD may be the most valuable predictors.

BACKGROUND:Mesenchymal stem cel s are the seed cel s for tendon tissue engineering which can be obtained in large quantities, but how to induce in vitro is a key technology.
OBJECTIVE:To explore the effect of platelet-rich plasma on the proliferation and col agen production of in vitro cultured mesenchymal stem cel s.
METHODS:The rabbit mesenchymal stem cel s were separated ad cultured. The high-dose platelet-rich plasma group, middle-dose platelet-rich plasma group and low-dose platelet-rich plasma group were set to induce the mesenchymal stem cel s, and the blank control group was set as control.
RESULTS AND CONCLUSION:The proliferation of mesenchymal stem cel s in the high-dose platelet-rich plasma group, middle-dose platelet-rich plasma group and low-dose platelet-rich plasma group was high, and in rapid growth with big increase amplitude, and there was no significant difference in the proliferation when compared with the blank control group (P<0.05). The effect was positively correlated with the culture time, and after cultured for a certain time, the effect was in dose-dependent manner, as higher dose platelet-rich plasma had more significant effect on the proliferation of the cel s. The results indicate that platelet-rich plasma can significantly promote the synthesis of col agen type Ⅰ and Ⅲ of mesenchymal stem cel s, the higher the dose, the more significant the effect on the col agen.

Abstract?AlM:To discuss the feasibility of using the visual quality analyzer KR - 1W to guide the relatively personalized aspheric intraocular lens ( lOL ) implants to make the whole eye spherical aberration close to 0. 1μm.?METHODS: ln this prospective case series study, the corneal spherical aberration with 6mm aperture of 73 patients (100 eyes) was measured with KR-1W Visual Function Analyzer 1d before surgery. For the sake of the whole postoperative spherical aberration were close to 0. 1μm, 9 cases ( 16 eyes ) with corneal spherical aberration <0. 15μm were implanted Sofport Advanced Optic lOL, named AO group;45 cases ( 57 eyes ) with corneal spherical aberration 0. 25~0. 3μm were implanted AcrySof lQ lOL, named lQ group;19 cases (27 eyes) with corneal spherical aberration > 0. 35μm were implanted Tecnis ZA9003 lOL, named Tecnis group. Aspherical lOL was implanted after phacoemulsification through a cornea 2. 75mm incision without suture. Uncorrected visual acuity, beat corrected visual acuity, spherical aberration of the whole eye and jnternal optics (mainly lOL) at 6mm pupil diameter were examined at 3mo postoperatively. The relevant data were analyzed using t-test and variance analysis.?RESULTS: The whole ocular spherical aberration at 6mm pupil diameter in all postoperative were 0. 084 ± 0. 032μm;in Tecnis group, the data were 0. 091 ± 0. 021μm;in AO group, the data were 0. 0814-0. 013μm;lQ group were0. 093 ± 0. 042μm. There was no significantly different between the predicted value and actual value of ocular spherical aberration at 6 mm pupil diameter in all postoperative ( t = 1. 932, P = 0. 061 ) and in the three groups. The difference value in the predicted values of the preoperative spherical aberrations of the whole eye and the actual values after surgery was 0. 013±0. 041μm; there was no statistically significant difference ( F=2. 537, P=0. 091 ) . Respectively compared the uncorrected visual acuity and besta corrected visual acuity among three groups of postoperative, no significant difference were found (F=0. 897, P=0. 421;F=1. 423, P=0. 097).?CONCLUSlON: Personality selection of aspheric lOL based on preoperative corneal spherical aberration of patients is feasible and produces satisfactory target postoperative total spherical aberration.

Objecfive To investigate the association between the exon 2,3,4 of MHC class Ⅰ chain-related gene-B(MICB)and ulcerative colitis(UC)in Chinese Han.Methods Using polymerase chain reaction single-stranded conformation polymorphism,allele frequency of MICB exon 2,3 and 4 in 105 patients with UC and 213 age and sex matched healthy controls were genotyped.All of the studied individuals were Chinese Han.Results Allele frequency of MICB 0106 was increased in patients with UC as compared with normal controls(19.0%vs 8.9%,P=0.000,Pc＜0.001,OR=2.402,95%CI:1.488-3.879).The frequency of MICB 0106 was increased significantly in patients with extensive colitis (24.4%vs 8.9%,P=0.000,Pc＜0.001,OR=3.294,95%CI:1.800-6.027),moderate and severe disease(24.1%vs 8.9%,P=0.000.Pc＜0.001,OR=3.294 95%CI:1.893-5.576)and in those with extra intestinal manifestations(20.5%vs 8.9%,P=0.002,Pc=0.012,OR=2.626,95%CI:1.418-4.861).Furthermore,MICB 0106 allele was higher in frequency in the male patients with UC (22.1%vs 8.0%,P=0.001,Pc=0.006,OR=3.276,95%CI:1.737-6.178)and the patients more than 40 years old(28.8%vs 8.3%,P=0.000,Pc＜0.001,OR=4.500,95%CI:2.381-8.504)as compared with healthy controls.Conclusion MICB 0106 allele is positively associated with UC,especially with extensive colitis,moderate and severe disease,presence of extra intestinal manifestations,male gender and age of more than 40 years in Chinese Han in Hubei province.

OBJECTIVETo evaluate the therapeutic effect on pain in the dressing change after perianal abscess surgery treated with warm suspending moxibustion at Changqiang (GV 1).

METHODSOne hundred and sixty patients were randomized into a moxibustion group and a far-infrared therapy group, 80 cases in each one. In the moxibustion group, the warm suspending moxibustion at Changqiang (GV 1) was applied. In the far-infrared therapy group, the infrared ray was radiated at Changqiang (GV 1). The treatment started in 10 min after dressing change in the two groups. Totally, 8 treatments were required. The time requirement being equal to or less than 2 score in the assessment face scale (AFS), the case number of pain relief in 10 min after treatment and the intention of the two therapies after everyday dressing change were observed in the patients of the two groups.

RESULTSIn the moxibustion group, the time of pain relief was shorter than that of the far-infrared therapy group [(11.5 +/- 3.9) min vs (17.8 +/- 3.8) min on the 1st day; (9.4 +/- 3.6) min vs (15.2 +/- 4.3) min on the 2nd day; (7.8 +/- 2.7) min vs (14.3 +/- 3.2) min on the 3rd day; (6.4 +/- 3.3) min vs (10.5 +/- 2.9) min on the 4th day; (5.9 +/- 2.9) min vs (11.2 +/- 1.9) min on the 5th day; (5.3 +/- 2.2) min vs (8.8 +/- 2.3) min on the 6th day; (5.4 +/- 1.8) min vs (9.2 +/- 2.1) min on the 7th day; (5.1 +/- 1.4) min vs (9.5 +/- 2.6) min on the 8th day, all P < 0.05]. The case numbers of pain relief in 10 min after treatment were 74 cases, 75 cases, 77 cases and 78 cases from the 1st to the 4th day separately in the moxibustion group, which were much more than 63 cases, 65 cases, 68 cases and 69 cases in the far-infrared therapy group (all P < 0.05). The treatment intention scores in the moxibustion group were better than those in the far-infrared therapy group (all P < 0.05).

CONCLUSIONThe warm suspending moxibustion at Changqiang (GV 1) relieves pain rapidly in the dressing change of much more patients after perianal abscess surgery in much shorter time as compared with the far-infrared therapy. Moreover, the patients have more expectation to be treated with moxibustion therapy.

Abscess ; surgery ; Adolescent ; Adult ; Aged ; Anus Diseases ; surgery ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Pain, Postoperative ; therapy ; Young Adult

ObjectiveTo explore the values of 3D enhanced T2-star weighted angiography (ESWAN) in detecting iron deposition of the brain in patients with multiple sclerosis ( MS).Methods Conventional MRI and 3D ESWAN were performed in 22 patients with released-remitting MS(RR-MS) and in 22 age- and gender-matched normal controls.Both the magnitude images and phase images were available after post processing of the original ESWAN data post processing.The expanded disability status scale (EDSS) scores of the patients were from 0 to 8.5,and the course of disease were between 0.5 year and 15 years.The manifestation of MS lesions was evaluated on conventional MR images and ESWAN images by two experienced radiologists with blind methods.The phase values were measured for MS lesions between the semioval center and periventricular white matter and the corresponding regions in the normal control group were measured as well The Wilcoxon rank test was used to compare the differences of phase values between MS patients and normal controls,and the Spearman rank correlation analysis were used to analyze the correlations among the phase values of MS lesions,EDSS scores and course of disease of MS patients.Results( 1 ) MS lesions were observed on magnitude images and phase images,respectively and together.Two hundred and thirteen(32.8％ )lesions were detected on both the magnitude images and phase images,164 (25.2％)lesions were detected on the magnitude images only,and 273 (42.0％) lesions were detected on the phase images only.A total of 650 lesions were observed on the ESWAN,which showed more 42 lesions than conventional images(608).Among 486(273 + 213 )lesions observed on the phase images,205 (31.5％) lesions were homogeneously hypointense,45 (6.9％ ) lesions showed hypointense ring,and 236 (36.4％ ) lesions were inhomogeneously hypointense.(2) “ Penetrating veins” were showed in 424 (65.2％ ) periventricular lesions.The “penetrating veins” showed dilated and prolonged in 198 (30.5％ ) acute lesions,thin and short in 208 (32.9％ ) chronic lesions.However,the “penetrating veins” of 12 ( 1.8％ ) chronic lesions in 5 stable MS patients were expanded and prolonged slightly.( 3 ) The average phase value of the lesions ( - 0.05 ± 0.08) in patients with RR-MS was significantly lower than that of corresponding to regions (0.06 ± 0.05 ) in the normal control group( Z = 22.30,P ＜ 0.01 ).(4) The phase values of MS lesions were significantly correlated to the EDSS and course of disease of patients with RR-MS (r = -0.98,0.99 respectively,P ＜ 0.01 ).ConclusionESWAN can be helpful in the evaluation and quantification of abnormal iron deposition in MS lesions,which provides important values for the pathogenesis and pathological study for MS patients.

Objective To investigate the value of PET-CT in TNM staging and three-conformal radiotherapy (3DCRT) in esophageal carcinoma. Methods From September 2007 to November 2008, 20 patients with pathologically confirmed esophageal carcinoma were enrolled, including 2 treated with surgery and 18 with 3DCRT. All the patients received PET-CT simulation before the treatment. The length and maximum transverse diameter of Grit based on esophagoscope, esophagography, CT, PET-CT_(SUV2.5) and PET-CT_(40%SUVmax) were compared. The TNM stages were compared based on CT and PET-CT images. Three treatment plans were produced and analyzed based on images of CT, PET-CT_(SUV2.5) and PET-CT_(40%SUVmax),respectively. Results The length of lesion on esophagoscope, esophagography, CT, PET-CT_(SUV2.5) and PET-CT_(40%SUVmax) was 4. 93 cm, 5.06 cm, 6. 67 cm, 5. 89 cm and 4. 84 cm, respectively. The corresponding maximum transverse diameter on the last 3 images was 4. 05 cm, 3.38 cm and 2. 95 cm, respectively. In all, 31, 21 and 14 positive lymph nodes were identified according to CT images, PET-CT images and the both. Five patients with CT diagnosed stage M_0 were found to have distant metastasis by PET-CT images, and 1 patient with CT diagnosed stage M_1 was excluded by PET-CT. The volumes of GTV_(CT) were similar with GTV_(SUV2.5) in 2 patients, smaller in 5 patients, and larger for the remaining 13 patients. For these 13 patients, the radiation dose of normal tissues based on GTV_(SUV2.5) was relatively lower. Conclusions The length of lesion based on PET -CT_(SUV2.5) matches the pathological length best , followed by esophagography. With PET-CT_(40%SUVmax) the actual lesion length may be underestimated. TNM stage might be changed by PET-CT, and then the target volumes and radiation doses of normal tissues might be reduced.