[Study on the assignment method of intensity modulated radiotherapy plan for nasopharyngeal carcinoma based on MR images].

Objective: To discuss the influence of different computed tomography (CT) value assignment methods on dose calculation of intensity modulated radiotherapy (IMRT) plan which designed for nasopharyngeal carcinoma (NPC) and the value assignment methods of IMRT plan for NPC based on magnetic resonance (MR) images. Methods: Simulation CT and MR image of 32 NPC patients in Shandong Cancer Hospital from March 2018 to November 2018 were selected for this study. Populate CT values were obtained by contouring and analyzing the simulation CT of patients' tissue, including bone, air, brain, eyeball, optic-nerve, lens, parotid, masseter, skin. Pseudo-CT were generated by different CT value assignment methods: CT1: CT value of all tissues was set to 0HU; CT2: CT value of air cavity was set to populate CT value based on CT1; CT3: CT value of Bone was set to populate CT value based on CT2; CT4: CT value of each soft tissue were set to populate CT value based on CT3. The IMRT plan for NPC as Plan0 was designed base on simulation CT. Then Plan0 was transplanted to four pseudo-CT to recalculate the dose and obtain Plan1, Plan2, Plan3 and Plan4, the differences of dosimetric parameters were compared with Plan0. NPC-IMRT plan was designed base on MR images by using the assignment method with CT value of each tissue were set to populate CT value. Results: In the head and neck CT images, the average populate CT values of bone and cavity were 621 HU and -720 HU, respectively. The populate CT values of other soft tissue ranges from -20 HU to 70 HU. The differences of dosimetric indexes of Plan1, Plan2, Plan3, Plan4 decreased sequentially compare to Plan0, the difference of the dosimetry parameters of Plan4 and Plan0 was the smallest. The differences of PTV D(99), PTV D(95), isocenter dose, D(mean) of all tissues, D(max) of bilateral eye balls, D(max) of bilateral lens, D(max) of bilateral optic nerves, D(mean) of bilateral parotid, V(20) of bilateral parotid, D(50) of bilateral parotid, D(max) of spinal cord, D(max) of brainstem, D(5) of brainstem between Plan4 and Plan0 were all less than 1%. The difference of V(30) in bilateral parotid between Plan4 and Plan0 was less than 1.5%. In the comparison of the pixel dose distribution, the regions of dose distribution difference greater than 1% mainly distributed in the air cavity, bone periphery and the skin. The target area of the IMRT plan for NPC based on MR images met 95% of the prescribed dose, and the dose of each organ at risk was within the dose limit. Conclusions: The assignment method of each tissue and organs set to populate CT value compared with other methods has the least influence on the dose calculation of NPC-IMRT plan, which could meet the clinical requirements. Therefore, it should be the first choice of assignment method when designing NPC-IMRT plan based on MR image.

[Application of radiomics captured from CT to predict the EGFR mutation status and TKIs therapeutic sensitivity of advanced lung adenocarcinoma].

Objective: To explore the ability of computed-tomography (CT) radiomic features to predict the Epidermal growth factor receptor (EGFR) mutation status and the therapeutic response of advanced lung adenocarcinoma to EGFR- Tyrosine kinase inhibitors (TKIs) treatment. Methods: A retrospective analysis was performed on 253 patients diagnosed as advanced lung adenocarcinoma, who underwent EGFR mutation detection, and those with EGFR sensitive mutation were treated with TKIs. Using the Lasso regression model and the 10 fold cross-validation method, the radiomic features of predicted EGFR mutation status and the screening of TKIs for sensitive populations were obtained. 715 radiomic features were extracted from unenhanced, arterial phase and venous phase, respectively. Results: The area under curve (AUC) values of the multi-phases including unenhanced, arterial phase and venous phase of the EGFR mutation status validation group were 0.763, 0.807 and 0.808, respectively. The number of radiomic features extracted from the multi-phases were 5, 18 and 23, respectively, which could distinguish the EGFR mutation status. The AUC values of the multi-phases of the EGFR-TKIs sensitive validation group were 0.730, 0.833 and 0.895, respectively. The number of radiomic features extracted from the multi-phases were 3, 7 and 22, respectively, which can be used to screen the superior population for TKIs treatment. The efficiency of radiomic features extracted from venous phase in predicting EGFR mutant status and EGFR-TKIs sensitivity was significantly superior than those of unenhanced and arterial phase. Conclusions: The radiomic features of CT scanning can be used as the radiomics biomarker to predict the EGFR mutation status of lung adenocarcinoma and to further screen the dominant population in TKIs therapy, which provides the basis for targeted therapy.

[HBsAg loss with Pegylated-interferon alfa-2a in hepatitis B patients with partial response to nucleos(t)-ide analog: new switch study].

Objective: Hepatitis B surface antigen (HBsAg) loss is seldom achieved with nucleos(t)ide analog (NA) therapy in chronic hepatitis B patients but may be enhanced by switching to finite pegylated-interferon (Peg-IFN) alfa-2a. We assessed HBsAg loss with 48- and 96-week Peg-IFN alfa-2a in chronic hepatitis B patients with partial response to a previous NA. Methods: Hepatitis B e antigen (HBeAg)-positive patients who achieved HBeAg loss and hepatitis B virus DNA < 200 IU/mL with previous adefovir, lamivudine or entecavir treatment were randomized 1:1 to receive Peg-IFN alfa-2a for 48 (n = 153) or 96 weeks (n = 150). The primary endpoint of this study was HBsAg loss at end of treatment. The ClinicalTrials.gov identifier is NCT01464281. Results: At the end of 48 and 96 weeks' treatment, 14.4% (22/153) and 20.7% (31/150) of patients, respectively, who switched from NA to Peg-IFN alfa-2a cleared HBsAg. Rates were similar irrespective of prior NA or baseline HBeAg seroconversion. Among those who cleared HBsAg by the end of 48 and 96 weeks' treatment, 77.8% (14/18) and 71.4% (20/28), respectively, sustained HBsAg loss for a further 48 weeks. Baseline HBsAg < 1 500 IU/mL and week 24 HBsAg < 200 IU/mL were associated with the highest rates of HBsAg loss at the end of both 48- and 96-week treatment (51.4% and 58.7%, respectively). Importantly, extending treatment from 48 to 96 weeks enabled 48.3% (14/29) more patients to achieve HBsAg loss. Conclusion: Patients on long-term NA who are unlikely to meet therapeutic goals can achieve high rates of HBsAg loss by switching to Peg-IFN alfa-2a. HBsAg loss rates may be improved for some patients by extending treatment from 48 to 96 weeks, although the differences in our study cohort were not statistically significant. Baseline and on-treatment HBsAg may predict HBsAg loss with Peg-IFN alfa-2a.

[Clinical effect and safety of pegylated interferon-α-2b injection (Y shape, 40 kD) in treatment of HBeAg-positive chronic hepatitis B patients].

Objective: To investigate the clinical effect and safety of long-acting pegylated interferon-α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 µg/week) in the treatment of HBeAg-positive chronic hepatitis B (CHB) patients, with standard-dose Peg-IFN-α-2a as positive control. Methods: This study was a multicenter, randomized, open-label, and positive-controlled phase III clinical trial. Eligible HBeAg-positive CHB patients were screened out and randomized to Peg-IFN-α-2b (Y shape, 40 kD) trial group and Peg-IFN-α-2a control group at a ratio of 2:1. The course of treatment was 48 weeks and the patients were followed up for 24 weeks after drug withdrawal. Plasma samples were collected at screening, baseline, and 12, 24, 36, 48, 60, and 72 weeks for centralized detection. COBAS® Ampliprep/COBAS® TaqMan® HBV Test was used to measure HBV DNA level by quantitative real-time PCR. Electrochemiluminescence immunoassay with Elecsys kit was used to measure HBV markers (HBsAg, anti-HBs, HBeAg, anti-HBe). Adverse events were recorded in detail. The primary outcome measure was HBeAg seroconversion rate after the 24-week follow-up, and non-inferiority was also tested. The difference in HBeAg seroconversion rate after treatment between the trial group and the control group and two-sided confidence interval (CI) were calculated, and non-inferiority was demonstrated if the lower limit of 95% CI was > -10%. The t-test, chi-square test, or rank sum test was used according to the types and features of data. Results: A total of 855 HBeAg-positive CHB patients were enrolled and 820 of them received treatment (538 in the trial group and 282 in the control group). The data of the full analysis set showed that HBeAg seroconversion rate at week 72 was 27.32% in the trial group and 22.70% in the control group with a rate difference of 4.63% (95% CI -1.54% to 10.80%, P = 0.1493). The data of the per-protocol set showed that HBeAg seroconversion rate at week 72 was 30.75% in the trial group and 27.14% in the control group with a rate difference of 3.61% (95% CI -3.87% to 11.09%, P = 0.3436). 95% CI met the non-inferiority criteria, and the trial group was non-inferior to the control group. The two groups had similar incidence rates of adverse events, serious adverse events, and common adverse events. Conclusion: In Peg-IFN-α regimen for HBeAg-positive CHB patients, the new drug Peg-IFN-α-2b (Y shape, 40 kD) has comparable effect and safety to the control drug Peg-IFN-α-2a.

[Efficacy and safety of pegylated interferon α-2b injection (Y shape, 40 kD) in treatment of patients with genotype 1/6 chronic hepatitis C].

Objective: To investigate the efficacy and safety of the new investigational drug pegylated interferon α-2b (Peg-IFN-α-2b) (Y shape, 40 kD) injection (180 µg/week) combined with ribavirin in the treatment of patients with genotype 1/6 chronic hepatitis C (CHC), with standard-dose Peg-IFN-α-2a combined with ribavirin as a positive control. Methods: A multicenter, randomized, open-label, and positive-controlled phase III clinical trial was performed. Eligible patients with genotype 1/6 CHC were screened out and randomly divided into Peg-IFN-α-2b(Y shape, 40kD) group and Peg-IFN-α-2a group at a ratio of 2:1. The patients in both groups were given oral ribavirin for 48 weeks in addition and then followed up for 24 weeks after drug withdrawal. Abbott Real Time HCV Genotype II was used to determine HCV genotype, and Cobas TaqMan quantitative real-time PCR was used to measure HCV RNA level at 0, 4, 12, 24, 48, and 72 weeks. Adverse events were recorded in detail. The primary efficacy endpoint was sustained virological response (SVR), and a non-inferiority test was also performed. Results: A total of 561 patients with genotype 1/6 CHC were enrolled, among whom 529 received treatment; 90.9% of these patients had genotype 1 CHC. The data of the full analysis set showed that SVR rate was 69.80% (95% CI 65.00%-74.60%) in the trial group and 74.16% (95% CI 67.73%-80.59%) in the control group (P = 0.297 0). The data of the per protocol set (PPS) showed that SVR rate was 80.63% (95% CI 76.04%-85.23%) in the trial group and 81.33% (95% CI 75.10%-87.57%) in the control group (P = 0.849 8), and the 95% CI of rate difference conformed to the non-inferiority standard. The analysis of the PPS population showed that of all subjects, 47.9% achieved rapid virologic response, with a positive predictive value of 93.8%. The incidence rate of adverse events was 96.30% in the trial group and 94.94% in the control group, and the incidence rate of serious adverse events was 5.13% in the trail group and 5.06% in the control group. Conclusion: In the regimen of Peg-IFN-α combined with ribavirin for the treatment of genotype 1/6 CHC, the new investigational drug Peg-IFN-α-2b(Y shape, 40 kD) has comparable clinical effect and safety to the control drug Peg-IFN-α-2a.

Comparison of the antiviral effects of entecavir and adefovir dipivoxil in chronic HBV infection: a randomized control trial.

AIMS: The purpose of this study is to compare the antiviral efficacy of entecavir (ETV) and adefovir dipivoxil (ADV) at various time points during the treatment. METHODS: A randomized, controlled, open-label study was designed to analyze the kinetics of HBeAg seroconversion, HBV DNA level, and liver and renal functions in 72 ETV-treated chronic hepatitis B (CHB) patients and 66 ADV-treated CHB patients. The data was collected every 12 weeks up to 96 weeks after drug administration. RESULTS: The negative rate of HBeAg seroconversion was significantly increased at 24 weeks in ETV-treated patients, whereas in ADV-treated patients, these changes were not significant. The serum HBV DNA levels were significantly decreased from 24 weeks in both ETV- and ADV-treated patients. Other than ETV showing significantly decreased levels of HBV DNA at 24 weeks when compared with ADV, there was no difference in virological response between two treatments at any other time points. The serum alanine aminotransferase (ALT) and total bilirubin (TBIL) levels were significantly decreased 12 weeks after either ETV- or ADV-treated patients without differences between two treatments. The urea nitrogen levels were in normal range and there was no difference between two groups. CONCLUSIONS: Our study suggested that both ETV and ADV could be used as monotherapy for nucleotide-naive patients, but ETV has displayed potential efficacy in HBeAg seroconversion.

RapidArc combined with the active breathing coordinator provides an effective and accurate approach for the radiotherapy of hepatocellular carcinoma.

PURPOSE: The goal of this research was to investigate the feasibility of volumetric modulated arc therapy, RapidArc (RA), in association with the active breathing coordinator (ABC) for the treatment of hepatocellular carcinoma (HCC) with radiotherapy. PATIENTS AND MATERIALS: A total of 12 patients with HCC, after receiving transcatheter arterial chemoembolization (TACE) treatment, underwent three-dimensional computer tomography (3D-CT) scanning associated with ABC using end inspiration hold (EIH), end expiration hold (EEH), and free breathing (FB) techniques. The three-dimensional conformal radiotherapy (3D-CRT), intensity-modulated radiotherapy (IMRT), and RA plans (three 135° arcs) were designed on different CT images, respectively. The liver volume, gross tumor volume (GTV), and planning target volume (PTV) of the three breath status and the dosimetric differences of the different plans were compared. RESULTS: There were no significant differences in the volumes of live and GTV between the three breathing techniques (p > 0.05); the PTV in FB was greater than in the EEH and EIH (p < 0.05). The overall conformality index (CI) and homogeneity index (HI) for RA (CI 0.92, HI 0.90) were better than IMRT (CI 0.90, HI 0.89) and 3D-CRT (CI 0.70, HI 0.84) for the three breathing techniques (p< 0.05). The RA and IMRT significantly reduced the mean dose, V(20), V(30), and V(40) of normal liver compared to 3D-CRT, while the V(5) and V(10) in RA were higher than in IMRT. The mean values in mean dose, V(10), V(20), V(30), and V(40) of the normal liver were reduced from 13.12 Gy, 46%, 24%, 13%, and 8% in RA(FB) to 10.23 Gy, 35%, 16%, 8%, and 5% in RA(EEH) and 9.23 Gy, 32%, 16%, 8%, and 5% in RA(EIH ), respectively. In addition, the treatment time of RA was equal to 3D-CRT, which was significantly shorter than IMRT. CONCLUSION: RA in conjunction with ABC for the treatment of HCC with radiotherapy can achieve better dose delivery and ensure the accuracy of the target volume, which spares more organs at risk, uses fewer monitor units, and shortens treatment time.

Molecular cloning of growth hormone-encoding cDNA of an Indian major carp, Labeo rohita, and its expression in Escherichia coli and zebrafish.

The cDNA clones encoding for growth hormone (GH) of an Indian major carp rohu Labeo rohita were isolated from a cDNA library constructed from the poly(A)(+) RNA extracted from the pituitary glands of rohu. Partial GH cDNA of the rohu (3'-end) was amplified by RT-PCR and used as probe to screen the cDNA library. Full-length GH-specific cDNA clones (1180 bp) were isolated and sequenced. The sequence contains 48-bp 5'-noncoding region followed by an ORF of 621 bp and a 3'-noncoding region of 521 bp. The peptide shares about 90% identity with the GH of Cyprinus carpio (Linn.) and >84% identity with GH sequences of other cyprinids. The GH-encoding cDNA of rohu has been cloned into expression vectors and GH protein has been over expressed in Escherichia coli and purified as a soluble protein. The GH cDNA was cloned into a bicistronic vector with EGFP; injection of in vitro transcribed GH-EGFP mRNA into zebrafish embryo has resulted in EGFP expression confirming the cloned GH cDNA is functional in fish and the IRES element could be effectively used in fish for bicistronic expression of foreign genes.

[Primary study of HCV replication in HepG2 cells inoculated by anti-HCV positive and HCVRNA RT-PCR positive serum].

OBJECTIVE: To study HCV replication in HepG2 cell line and to establish a stable HepG2 cell line infected with HCV. METHODS: Continuous inoculation of anti-HVC positive and HCVRNA RT-PCR positive sera was used to infect HepG2 cells. The plus and minus strains of HCVRNA were detected by reverse transcription polymerase chain reaction in the HepG2 cells. Virus particles were observed in postinoculated HepG2 cells by transmission electron microscope. Immunohistochemical staining was used respectively to detect HCV antigen, HCVNS5 and HVC capsid in postinoculated HepG2 cells. RESULTS: HCV RNAs (both plus and minus strains) were found from the first passage to the third passage of postinoculation HepG2 cells, and HCV viruslike particles were detected in the third passage of the infected HepG2 cells, those particles mostly occurred within cytoplasm, their diameters were between 30-60 nm, and gathering silver particles were found in immunogold silver staining of HCV NS5/capsid protein in infected HepG2 cells. Gathering gold particles were found mostly within vesicles in cytoplasm by immuno gold electron microscopy. CONCLUSION: HepG2 cell line infected by HCV is established by continuous inoculation of anti-HCV positive and HCVRNA positive sera. HCV can replicate and produce virus-like particles in HepG2 cells, which may occur in the serial passages of post-infected HepG2 cells.

[Relationship between hot spot mutation in hepatitis B virus basic core promotor and HBeAg status].

OBJECTIVE: This paper was to study the influence of hot spot mutation in hepatitis B virus (HBV) basic core promotor (BCP) (nt1762 and nt1764) on HBeAg status of asymptomatic HBV carriers. METHODS: Mismatched PCR was used to amplify the fragment of HBVBCP and the fragments were analysed by restrict enzyme assay. Ninety cases of HBV infection were tested for hot spot mutations in HBVBCP. RESULTS: Twenty-six (43.3%) of 60 asymptomatic HBV carriers with HBeAg negative were found to have hot spot mutations in HBVBCP, among which 20 cases were accompanied by the mutation of nt 1896 in HBV Pre-C region. Hot-spot mutation in HBVBCP, however, occurred only in 3(10%) of 30 asymptomatic HBV carriers with HBeAg positive. For further investigation the HBVBCP hot spot mutations in asymptomatic HBV carriers without mutation of HBV Pre-C region were studied. Six(31.6%) of 19 asymptomatic HBV carriers with HBeAg negative were found to have hot spot mutation in HBVBCP. Two(7.1%) of 28 asymptomatic HBV carriers with HBeAg positive had hot spot mutations in HBVBCP, and there was a statistically significant difference between two groups. CONCLUSIONS: Hot spot mutations in HBVBCP is common in the HBV carriers with HBeAg negative and usually accompanies with nt1896 mutation of HBV Pre-C region. It is possible that Hot spot mutation in HBVBCP is a new reason of HBeAg negative HBV infection.