Objective To evaluate the efficacy and drug resistance profiles of nucleosides (NA) retreatment in NA experienced chronic hepatitis B (CHB) patients. Methods Totally 104 NA experienced CHB subjects were enrolled in this study.All these subjects had received at least 3 months NA monotherapy and stopped the treatment,and then received NA retreatment for at least one year.The subjects were divided into three groups according to the following criteria:reached the therapy endpoint of China guideline when they stopped NA-naive treatment (group A,n =39); did not reach the therapy endpoint when they stopped NA-naive treatment but hepatitis B virus (HBV) DNA＜1.0× 103 copy/mL (group B,n=33); and with HBV DNA＞1.0× 103 copy/mL (group C,n=32).The efficacy and drug resistance profiles of retreatment were compared among three groups. The effects of baseline alanine aminotransferase (ALT) levels,HBV DNA levels and HBeAg titers on the retreatment efficacies were analyzed. The mutations of HBV P gene were detected by nested polymerase chain reaction (PCR) and direct sequencing.The data were analyzd by Wilcoxon test and x2 test.Results The time to ALT normalization in patients with baseline ALT＜ 1.3 × upper limit normal (ULN) was shorter than that in patients with ALT≥1.3×ULN (2 months vs 4 months; Z=2.281,P=0.023).The time to virological response in patients with baseline HBV DNA＜5 lg copy/mL was shorter than that in patients with HBV DNA≥5 lg copy/mL (1 month vs 2 months; Z=2.054,P =0.040). The time to virological response and ALT normalization in baseline HBeAg negative were both shorter than those in patients with baseline HBeAg positive patents ( 1 month vs 3 months and 2 months vs 4.5 months,respectively; Z=2.580 and 2.304,respectively; both P＜0.05). The subjects in group A achieved virological response and HBeAg seroconversion after retreatment earlier compared to previous NA-naive therapy ([1.61 ± 1.76] months vs [3.48±4.066]months and [3.38 ± 3.34] months vs [9.92-11.22] months,respectively; Z=-2.854 and-1.094,respectively; both P＜0.05).The cumulative HBeAg seroconversion rate in group A was higher compared to those in group B and group C (80.0％ vs 36.8％ and 37.5％,respectively; x2 =4.368 and 5.100,respectively; both P＜0.05).Thirteen patients developed clinical resistance and four of them developed genotypic resistance proved by PCR direct sequencing.Among the patients retreated with the same regimen as previous in the C group,the cumulative resistance rate was highest compared to group A and B (44％ vs 9％ and 0,respectively; x2 =5.019 and 6.588,respectively;both P＜0.05).No resistance was detected in the 14 patients retreated with combined NA treatment without cross resistance.Conclusions For NA experienced CHB patients who fulfill the indication of antiviral therapy,the retreatment should be started as earlier as possible. Retreatment with NA combination without cross resistance can prevent (reduce) the risk of developing drug resistance.

Objective To explore the influence factors on hepatitis B virus (HBV) relapse after nucleos(t)ide analogues (NA) withdrawal in the chronic hepatitis B (CHB) patients who met NA cessation criteria. Methods Eighty-one consecutive CHB patients were treated with NA, 38 with lamivudine (LAM), 25 with adefovir dipivoxil (ADV), 12 with entecavir (ETV), 6 with LAM +ADV. Among recruited patients, 40 were hepatitis B virus e antigen (HBeAg) positive, 41 were HBeAg negative, 67 of them were initial treatment, 14 were retreatment due to resistance to NA at baseline. The treatment was discontinued after meeting China therapeutic end-point criteria. HBV DNA, HBV serological markers, alanine aminotransferase (ALT) were measured respectively at baseline, every month before virological response, every 3 months after virological response, every month within first 6 months and every 2 months over 6 months after drugs withdrawal. Twelve probable influence factors on relapse which were sex, age, HBV family history, baseline HBV DNA,baseline HBeAg status, baseline ALT, virological response time, total duration of treatment, duration of additional treatment, the level of hepatitis B virus surface antigen (HBsAg) at cessation therapy,initial treatment or retreatment, drug category were analyzed with univariate, multivariate Cox regression modle and stratified analysis. The cumulative relapse was calculated by the Kaplan-Meier method. Results A total of 36 patients (44. 4%) relapsed within 1 year. Initial treatment or retreatment, HBV family history, virological response time, the level of HBsAg at cessation therapy were independent risk factors. The relapse rate of retreatment was higher than that of initial treatment (78.6% vs 37. 3% , χ2 = 7. 983, P = 0. 005) , those of patients with HBV family history higher than without family history (64. 5% vs 15.0%, χ2 =12. 096,P = 0.002), those of patients obtained virological response within 3 months lower than after 3 months(34. 0% vs 64. 3% , χ2 =6. 823,P=0. 009) , those of patients with HBsAg≤150 μg/L at cessation therapy lower than >150 μg/L(27. 6% vs 53. 8%, χ2=5. 199,P=0. 023). Conclusions Retreatment, HBV family history, later virological response and higher HBsAg level at cessation therapy are risk factors of relapse after NA withdrawal. Such patients should be treated with prolonged duration after meeting end-point criteria to strengthen the efficacy.

This study was aimed to identify pET21b-HPV16E2/BL21(DE3) strain and to optimize the expression of human papillomavirus type 16 (HPV16) E2 protein by orthogonal analysis. Four influence factors on two levels were selected to increase the target protein quantity. The four factors were induction time, induction temperature, inductor concentration and cell density. The quantity of HPV16 E2 protein was used as the evaluation parameter. Induced by IPTG, HPV16 E2 protein was analyzed by SDS-PAGE and Western Blot. Target protein was analyzed by GIS imaging system to quantify the protein level. SPSS13. 0 software was applied to analyze the result. Data showed that the expression strain pET211rHPV16 E2/BL21(DE3) was identified correctly. HPV16 E2 protein expressed mainly at insoluble form. The 42KD protein band was identified by SDS-PAGE and Western blot. Orthogonal test was applied on influence factor analysis and expression optimization successfully. Main influence factors were inductor concentration and induction temperature. The optimimum condition of maximum expression quantity was 37 degrees C, 7h, 1.0 mmol/L IPTG and OD600 1.0. In this experiment, orthogonal test could not only be used to analyze the influential factors and promote the target protein expression, but also be used to provide a better experiment method for molecular biological study.
DNA-Binding Proteins ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Human papillomavirus 16 ; metabolism ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomavirus Infections ; virology ; Recombinant Proteins ; biosynthesis ; genetics

Objective To study the optical imaging quality of multifocal intraocular lens (IOL) following Nd∶YAG laser damage.Methods Three hydrophobic acrylate discs used to make multifocal IOL were placed in line,and the Nd∶YAG laser was used to emit the posterior surface,mid-plane and anterior surface of the discs by gradually increasing the emitting energy to determine the damaged threshold energy of various planes.Eight multifocal IOLs were divided into 2 groups.IOL damage models were established by emitting 12 points along 12 clocks on circle of 3.0 mm in diameter in the group 1 (IOLs NO.1,2,3,4) and 6 points with 1-clock interval on circle of 1.0 mm in diameter in the group 2 (IOLs NO.5,6,7,8) with Nd∶YAG laser.The modulation transfer function (MTF),line spread function (LSF) and United States air force (USAF) test target images of the IOLs were tested under the optical aperture at 2.0,3.0 and 4.5 mm.Results The damage thresholds of laser power were not significantly different among three hydrophobic acrylate discs (Fgroup =1.100,P =0.337),and the thresholds were significantly different among different planes in the discs (Fposition =195.279,P=0.000),with the highest damage threshold in the anterior surface and the lowest damage threshold in the mid-plane of the discs(all at P＜0.01).The shifts of far focus and near focus MTF curves,LSF curves and USAF target images of the IOLs under the 2.0,3.0 and 4.5 mm optical aperture after laser damage were in coincidence with those before laser damage both in group 1 and group 2.The glare phenomenon of the USAF target image was observed at near focus under the 4.5 mm optical aperture both before and after laser damage.Conclusions Multifocal IOL is vulnerable to injury when the laser beam focuses on the midplane,and the anterior surface of IOL can tolerate certain degree of damage.In standard model eye,the MTF curves,LSF curves and USAF target images of multifocal IOL was not impacted by laser induced spots injuries under different pupil diameters.

ObjectiveTo clarify the spatial distribution characteristics of medicinal plant resources in Gansu province， analyze the causes， changing trends， and driving factors of the spatial differentiation， and thus lay a scientific basis for the rational development and sustainable development of medicinal plant resources in this province. MethodBased on the data of The Fourth National Survey of Chinese Medicine Resources， the richness and spatial distribution difference of medicinal plant resources in 87 counties （districts） of Gansu province were analyzed via the global spatial autocorrelation analysis， trend surface analysis， local spatial autocorrelation analysis， and hotspot analysis. Moreover， the correlation of vegetation type， soil texture， annual average temperature， annual average precipitation， and altitude with the spatial distribution pattern of the medicinal plant resources was discussed. ResultCounties （districts） with high or low richness of medicinal plant resources in Gansu province were respectively clustered together. To be specific， counties （districts） with high richness of the medicinal resources were mainly in southeastern Gansu， while those with low richness in northwestern Gansu. The leading driving factors affecting the cold and hot spots included vegetation type， soil texture， and average annual rainfall. ConclusionThe species richness of medicinal plant resources in Gansu province rises from west to east and from north to south. The natural driving factors are the key to the diversity and spatial distribution pattern of medicinal plant resources， which show significant influence on them.