Objective To investigate the biological characteritics of adult animal hepatocytes induced by phenobarbital sodium(PBS) in vivo,and to study the potential value of biological artificial liver of effective hepatocytes.Methods 12 adult male mice,are yandomly divided in to preinducing group and controll group.The preinducing group are intraperitoneally injected PBS per day,45mg/kg for 7 times in total;the controll ware injected NS.after that,we detected the blood serum TP,BUN,CHOL,HDLC.And the same amount of isolative hepatocytes was developed after being developed 48h;MTT was used to investigate the proliferation of hepatocytes after being developed 24h;chromosome was investigated to observe the cell division;and the survival deadline and morphology was also investigated.Results The TP had remarkable difference between the two groups(t=2.678,P

Objective To establish HIV-1 seroconversion panels with the samples collected by National Institutes for Food and Drug Control ( NIFDC) and to evaluate the window periods of HIV enzyme immunoassay ( EIA) diagnostic kits used for blood screening with them. Methods Serum specimens were collected from different plasma donation stations in China. All suspected HIV infection specimens were screened for HIV by using the nucleic acid amplification testing (NAT), Western blot confirmatory assay and P24 quantitative detection assay. The HIV env gene sequences were amplified by RT-PCR for further confirmation of HIV infection. The PCR products were sequenced and genotyped. The confirmed seroconver-sion panels were used to evaluate the early detection capabilities of the 4th and 3rd generation HIV EIA diag-nostic kits used in blood screening in china. Results A total of 8 sets of HIV seroconversion panels com-prised of 36 samples were confirmed in this study, including 6 sets of AE subtype, 1 set of B subtype and 1 set of unknown genotype. Those seroconversion panels were tested with HIV diagnostic kits produced by 19 different manufacturers. For the early detection of HIV infection, the 4th generation HIV diagnostic kits with a score of 9. 4 points were better than the 3rd generation HIV diagnostic kits whose score was 3. 6 points (P<0. 01, t=8. 547). Some of the domestic 4th generation HIV diagnostic kits were similar to the imported kits in the early detection of HIV infection. In terms of the diagnosis of HIV infection, the HIV-1 NAT was at least 2 weeks earlier than the HIV EIA diagnostic kits. The sensitivity of confirmatory assay was lower than that of the diagnostic kits. Four out of five 4th generation HIV diagnostic kits showed declined signal to cut off ( S/CO ) ratio , indicating the probability of false detection during the second window period . Conclusion Eight sets of NIFDC HIV-1 seroconversion panels were established in this study. With those panels we found that there were differences in the window period between different EIA diagnostic kits used for HIV blood screening.

Objective To compare cellular immune responses in mice elicited by Chinese different AIDS candidate vaccines.Methods According to their different immunization procedures,BALB/c mice were immunized with 6 AIDS candidate vaccines,separately.Spleen cells were isolated for the detection of cellular immune response to HIV-specific peptides using enzyme-linked immunosorbent spot(ELISPOT)assay and intracellular cytokine staining(ICS)method.Results AIDS vaccines were evaluated by using potential T-cell epitopes(PTE)Gag,Env and Pol peptides pool and ELISPOT.The positive conversion rates for cellular immune response of 1#-6# vaccines fluctuated from 70% to 100%.The vaccine-induced cellular immune responses to specific peptides pool are different not only in magnitude but also in breadth.The Th1type cytokines,IFN-γand IL-2,were detected with ELISPOT in 1# and 2# vaccines.The productions of IFN-γand IL-2 induced by both of the two vaccines showed a moderate correlation(r1 =0.62,P1 ＜0.01 ;r2=0.79,P2 ＜ 0.01).The positive conversion rate of IFN-γ secreting cells of 1 # vaccine was 66.7%(10/15)mice detected with both ELISPOT and ICS.And the results tested by ELISPOT and ICS showed moderate correlation(r = 0.55,P ＜ 0.05).Conclusion The magnitude and breadth of cellular immune responses induced by different AIDS candidate vaccines are different.Being induced by different AIDS candidate vaccines,the IFN-γand other Th1 type cytokines detected by ELISPOT or ICS could be used to evaluate the cellular immune responses in mice.

Objective To investigate the feasibility of low-c oncentration iso_osmolar contrast agent together with low tube voltage and iterative reconstruction algorithm in rabbit liver computed tonography (CT) perfusion imaging.Methods A total of 15 bealthy New Zealand rabbits were scanned twice of liver CT perfusion scans each with 24 hours interval.The first scan (routine group) was acquired at 100 kV and 100 mAs with ultravist (370 mg/ml),while the second (double low group) was acquired at 80 kV and 100 mAs with iodixanol (270 mg/ml) at 24 hours after the first scan.The obtained images were reconstructed with filtered back projection (FBP) and adaptive iterative dose reduction (AIDR-3D)algorithms in the controlled and experimental groups,respectively.The perfusion parameters including hepatic artery perfusion(HAP),portal vein perfasion(PVP),hepatic perfusion index(HPI),and total liver perfusion(TLP) and image quality as image quality score,average CT value of abdomen aorta,signalto-noise ratio(SNR),carrier-to-noise ratio(CNR),and figure of merit(FOM) were compared used pair ttest or Mann-Whitney U-test between the two groups wherever appropriate.The effective radiation dose and iodine intake were also recorded and compared.Results The image quality and perfusion parameters had no significantly different between the two groups except for FOM.The effective radiation dose and iodine intake were 38.79％ and 27.03％ lower in the double low group.Conclusions Low concentration iso _osmolar contrast agent (iodixanol,270 mg/ml) together with low tube voltage (80 kV) helps to reduce radiation dose and iodine intake without compromising perfusion parameters and image quality in liver CT perfusion imaging.

Objective To analyze the practicability of using ELISA kit for the detection of hepati-tis E virus antigen ( HEV-Ag) in plasma donations and Biomex HEV seroconversion panels. Methods The HEV-Ag positive samples were screened out from 36 340 donated blood plasma samples. Real-time fluores-cent PCR was performed for the detection of HEV RNA in HEV-Ag positive samples. The open reading frame 2 (ORF2) in HEV RNA was amplified by nested RT-PCR and the amplified products were confirmed by sequencing analysis. Phylogenetic tree was constructed for HEV genotyping. Five Biomex HEV serocon-version panels were used in this study for the detection of HEV-Ag, anti-HEV antibody and HEV RNA as well as the correlation analysis between HEV-Ag and HEV RNA. Results Twenty-six out of 36 340 plasma samples (0. 07%) were positive for HEV-Ag. Of the 26 samples, 25 samples were positive for HEV RNA as indicated by the results of nested RT-PCR and 23 positive samples were confirmed by sequencing analysis. The positive rate of HEV RNA in blood plasma donators was 1 ∶ 1 580 (0. 06%). There were 17 samples of genotype 1 (74%) and 6 samples of genotype 4 (26%) according to the phylogenetic tree analysis. All of the HEV-Ag positive samples were also positive for HEV RNA as indicated by the analysis of Biomex sero-conversion panels. HEV-Ag was consistent with the peak of the HEV RNA concentration. Conclusion A close relationship between HEV-Ag and HEV RNA was observed. HEV-Ag screening could be used as a measure to reduce the risk of HEV transmission by blood transfusion.

Objective To establish a high throughput phenotypic test for the detection of drug re-sistance in human immunodeficiency virus(HIV)strains. Methods The gene encoding luciferase was in-activated through restriction enzyme digestion and ligation. LacZ gene was used to replace the genes encoding original protease and reverse transcriptase. pol genes were amplified from pSG3△env plasmid and cloned in-to a new backbone plasmid through infusion. The factors that might affect the results of the test were opti-mized. Results The parental backbone plasmid pNL4-3. Lac was constructed,of which the gene encoding luciferase was inactivated and bearing the LacZ gene instead of genes encoding protease and reverse tran-scriptase. Several influential factors including cell numbers(10 000 / well),virus inoculation(200 TCID50 /well)and the concentration of DEAE-dextran(15 μg/ ml)were optimized. The reproducibility of this test was confirmed by testing 12 anti-HIV drugs against 2 pseudovirus strains 8 times,presenting the coefficient of variations(CVs)from 4. 32% to 28. 46% . Six types of pseudovirus were constructed and tested against the 12 anti-HIV drugs,the results of which were compared with those by using the pSG3△env-based pseud-ovirus test. The results of the two tests presented good consistency. Conclusion The high throughput phe-notypic test based on pNL4-3. Lac plasmid,combining the advantages of pSG3△env and pNL4-3 systems, could be used to analyze the drug resistance patterns of HIV-1 infectors and screen new drugs for antiretrovi-ral therapy in a rapid and effective way.

Objective To explore the feasibility of coronary computed tomographic angiography (CCTA) for obese patients with lower tube voltage (100 kV) and lower contrast media concentration (270 mgI/ml) using iterative reconstruction.Methods A total of 48 patients with body mass index greater than 30 kg/m2 were included and randomly divided into 2 groups according to random number table method.The images of the control group were obtained using iodine 370 mgI/ml, a tube voltage of 120 kV, and traditional filtered back projection (FBP) image reconstruction.Patients in the test group were injected with isotonic low concentration contrast media (270 mgI/ml), scanned with a lower tube voltage (100 kV), and adaptive iterative noise reduction image reconstruction algorithm (AIDR-3D) was used.Two experienced physicians scored the image quality in a double-blind way.Independent sample t-test was used to compare the effective dose (E), average CT values, signal to noise ratio (SNR), contrast to noise ratio (CNR), the figure of merit (FOM), image quality scores and the total iodine intake.Side effect was also evaluated.Results The subjective scores for control group and test group were not significantly different (P ＞ 0.05).The scores of two physicians were consistency (Kappa =0.88, P ＜ 0.05).The average CT values, SNR and CNR for the two groups were not significantly different (P ＞ 0.05), but the FOM of the test group was significantly higher than that of the control group (t =-9.250,-8.604,-9.158,-5.341, P ＜ 0.05).Effective dose in the test group was (1.61 ± 0.41) mSv, lower than that of the control group (t =8.373, P ＜ 0.01).The total iodine and iodine injection rate in the test group were both lower than in the control group (t =7.628, 8.480, P ＜ 0.01).The incidence of contrast mediarelated discomfort in the test group was lower than control group (x2 =18.70, 6.25, P ＜ 0.05).Conclusions For obese patients, isotonic low concentration of contrast media and low-dose CCTA could be feasible, which substantially reduce the radiation dose and iodine intake without sacrificing image quality.Trial registration Chinese clinical trial registry, ChiCTR-DPD-15007510.

Objective:To evaluate the effect of web-based problem-based learning software on the Clinical Medicine PBL Comprehensive Course. Methods:Thirty-eight students from the pilot class of undergraduate excellent doctors in clinical medicine were selected as the research objects. Based on the test scores of web-based problem-based learning software before the start of the course, the improvement of scores after passing the training of the Clinical Medicine PBL Comprehensive Course were compared, and the students' clinical thinking were quantitatively evaluated. Results:After all students completed the course, their abilities of expression and communication, teamwork and cooperation, problem analysis and clinical reasoning, and knowledge information management of the students increased steadily with the increase in the number of cases, but it was statistically significant after two PBL case trainings ( P<0.05). Learning intentions and attitudes scored the highest in the second PBL case (acute myocardial infarction), with statistical differences ( P<0.05). After training through the PBL course, diagnostic performance, clinical thinking, treatment scores, and overall performance scores of the students were significantly increased than before, with statistical significance ( P<0.05). The scores of case report writing and knowledge acquisition were not obviously improved. Conclusion:The combination of formative evaluation and web-based problem-based learning software can better evaluate the teaching effect and help students to understand their own shortcomings.

OBJECTIVE： To explore the mechanism of Lysimachia christinae in the treatment of cholecyst related diseases by network pharmacology. METHODS： The active ingredients of L. christinae were screened through TCMSP database with “Lipinski rule” and “Oral bioavailability ＞30％” rules， and their related targets were predicated correspondingly， then compound-target network were constructed by Cytoscape 3.2.1 software. Disease related targets were predicted by searching TTD database， OMIM database， PharmGKB database， DrugBank database and GAD database with “cholelithiasis” “gallstones” “cholecystitis” and “cholangitis” as keywords. Then， the network of disease-target was constructed and merged with active ingredient target to obtain therapeutic target. After pathway enrichment analysis of therapeutic target were performed by utilizing the DAVID database， molecular docking between target and active ingredient was also conducted in order to screen the main active ingredients of L. christinae. RESULTS： Twenty-seven active ingredients with good oral absorption and drug-like properties were screened from L. christinae. Thirty-three targets were attained after constructing and merging the network. Seven pathways， mainly related to cancer pathway and ABC transporter pathway were achieved. Top 4 active ingredients of L. christinae in the list of docking score were kaempferin， acacia， hesperetin and isorhamnetin， which acted on ABCC3， ABCB1， ABCC2， ABCB4 target. CONCLUSIONS： L. christinae treat cholecyst related diseases through ABC transporter pathway.

OBJECTIVES: Clinical research plays a vital role in disease research and population health. The public is the main source of clinical research volunteers. Understanding the public's cognition of clinical research plays a decisive role in the development of clinical research. This study aims to understand the Chinese public's cognition for clinical research and the influencing factors. METHODS: The questionnaire based on Chinese-translated Public Awareness of Research for Therapeutic Advancements through Knowledge and Empowerment (PARTAKE) was used to investigate the public's cognition for clinical research. RESULTS: Of the 2 513 valid respondents, 91.84% had heard of "clinical research", 91.76% of the respondents believed that clinical research was beneficial to society, 65.90% were willing to participate in clinical research, 87.50% believed that confidentiality was a very important thing, 73.70% believed that their personal information had been protected when participating in clinical research, and, 46.40% did not know whether volunteers participating in clinical research could receive adequate compensation. Educational levels, employment status, and annual income impacted in public perceptions of willingness to participate in clinical research, especially in privacy protection, informed consent, whether clinical research is intended for society, compensation for clinical research, and safety of clinical research (all P<0.01). CONCLUSIONS The Chinese public's cognition level for clinical research is acceptable, but there is still a lot of room for improvement in privacy protection, informed consent, and compensation. By designing a reasonable knowledge training program for clinical research and using the multimedia, improving access to the relevant knowledge, more public will know about clinical research recruitment information, which is of great significance for the development of clinical research in China.
Humans ; China ; East Asian People ; Educational Status ; Surveys and Questionnaires ; Public Opinion ; Knowledge ; Biomedical Research