Development and validation of ELISA method for detection of human signal regulatory protein α-anti-CD20 mouse chimeric antibody fusion protein IMM0306 in human serum / 药学学报

IMM0306 is a recombinant human signal regulatory protein α-anti-CD20 mouse chimeric antibody fusion protein, intended to treat refractory or recurrent CD20 positive B-cell non-Hodgkin lymphoma (B-NHL) in clinical. In this study, an ELISA method was established to evaluate the pharmacokinetics of IMM0306 in human serum. The experiment was approved by the Ethics Committee of the Cancer Hospital of the Chinese Academy of Medical Sciences (No. CTR20192612). Recombinant human CD47 protein was coated with the plate overnight, blocking the plate with 5% skin milk for 2 h. After washing, 100 μL per well standard and unknown samples were added, and incubated for 1.5 h. After washing, the detection antibody Anti-IMM0306-Biotin was added and incubated for 1 h, and then HRP-labeled streptavidin was added for 1 h, and the color was detected. The optimal concentration of coating reagent was 2 μg·mL-1 by ELISA method, and the optimal dilution of anti-IMM0306-biotin and SA-HRP were 1∶500 and 1∶5 000, respectively. The lower limit of quantitation was 4 ng·mL-1, and the standard curve range was 4-100 ng·mL-1. The verification results of the method meets the corresponding acceptance criteria, and can be used in IMM0306 clinical pharmacokinetic studies.

Research on the curative effect of standard channel PCNL combined with ultrasonic pneumatic lithotripsy in the treatment of upper urinary tract calculi under the guidance of ultrasound / 中国医学装备

Objective:To study the curative effect of standard channel (percutaneous nephrolithotomy) PCNL combined with ultrasonic pneumatic lithotripsy in the treatment of upper urinary tract calculi under the guidance of ultrasound, and analyze the influence factors of post-operative residual calculi.Methods: The data of 94 patients with upper urinary tract calculi were researched by using retrospective analysis. All of patients underwent the treatment of standard channel PCNL combined with ultrasonic pneumatic lithotripsy for upper urinary tract calculi under the guidance of ultrasound. The curative effects of upper ureteral calculi, pelvis calculi, multiple renal calculi, kidney stone with ureteral calculi and renal staghorn calculi were observed, and the post-operative complications were further detected so as to grasp the situation of residual calculi and calculate the clearance rate of calculus. The difference of general documents between patients with residual calculi and those without residual calculi were compared, and the influence factors of residual calculi were analyzed by using Logistic regression equation.Results:The incidence of post-operative complication was 27.66%, the clearance rate of calculus was 67.2% and residual rate of calculus was 32.98%. The differences of location puncture time, establishing channel time and ostomy fistula time among the five kinds of patients were no significant (F=1.89,F=2.46, F=0.91,P>0.05). While the differences of calculus removed time, amount of bleeding in intra-operative period and length of stay among various kinds of patients were significant (F=81.90,F=35.84,F=4.17,P<0.05). Depended on the results of Logistic regression equation, the types of calculus, size of calculus, hydronephrosis, urinary tract infection and renal insufficiency were the influence factors of post-operative residual calculus of patients with upper urinary tract calculi who received the treatment of PCNL combined with ultrasonic pneumatic lithotripsy.Conclusion: PCNL combined with ultrasound pneumatic lithotripsy under the guidance of B ultrasound without major complications has better curative effect, and its clearance rate of calculus is higher. Besides, the types of calculus, size of calculus, hydronephrosis, urinary tract infection and renal insufficiency can increase the incidence of post-operative residual calculus and affect the curative effect.

Prevalence of Parvovirus B19 Infection in Chinese Xiamen Area Blood Donors / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To estimate the prevalence of parvovirus B19 infection in Chinese Xiamen area blood donors.</p><p><b>METHODS</b>Blood samples from blood donors were tested for detection of parvovirus B19 DNA and antibody. The direct sequencing and genetype analysis of B19 DNA positive samples were performed.</p><p><b>RESULTS</b>Six out of 10452 samples were B19 DNA positive. The viral loads of the 6 samples were between 3.59×10-1.07×10IU/ml; the positive rate of B19-IgM was 4.64%(50/1078) and B19-IgG was 16.79%(181/1078). The positive rate of B19-IgG increased with ages, and was not related with the sex.</p><p><b>CONCLUSION</b>The overall prevalence of parvovirus B19 infection in blood donors is lower in Chinese Xiamen area than that in other areas, however, there is still a certain percentage of viremia in donors and the attention should be paid to blood safety in the future work.</p>

Epidemiologic Survey of Blood Donors with HBsAg⁻/HBV DNA⁺ in Chinese Xiamen Area / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To understand the characteristics of infections from blood donors with HBsAg⁻/HBV DNA⁺ in Xiamen area.</p><p><b>METHODS</b>Donors in Xiamen area were assayed by routine ELISA and those with negative results were tested by nucleic acid amplification testing (NAT). HBsAg⁻/HBV DNA⁺ samples were tested by quantitative detection of HBV DNA. Epidemiological analysis and following up examination were conducted in HBsAg⁻/HBV DNA⁺ donors.</p><p><b>RESULTS</b>Out of 130659 samples 113 were tested as HBsAg⁻/HBV DNA⁺ and with a rate of 0.09%. Among those, 62 samples were tested by quantitative detection of HBV DNA. All of the quantitative results were less than 1 × 10³ IU/ml and 93.5% (58/62) of which were less than 100 IU/ml. The possitive rate of HBsAg⁻/HBV DNA⁺ donors rose with ages. The possitive rate in male donors was higher than that in female and was lower in highly educated ones. Students and public servants had a lower positive rate.</p><p><b>CONCLUSION</b>The possitive rate of HBsAg⁻/HBV DNA⁺ donors is higher in Xiamen and the distribution of possitive donors has certain epidemiological characteristics. It is necessary to mobilize and recruit more people with a lower rate of HBsAg⁻/HBV DNA⁺ infection.</p>

Surveillance for occult HBV infection and HBsAg variants in blood donors / 病毒学报

Occult hepatitis B virus (HBV) infection status of blood donors in a southern city in China was investigated by immunological assays and nucleic acid testing. Overall, 17 (0.19%, 95% CI: 0.11%-0.30%) of the 9023 HBsAg negative samples were found to be positive for the presence of HBV DNA. "A" epitope sequences were obtained from 14 among them. Mutation(s) in aa124-aa147 existed in 6 (42.9%, 6/14) samples and 4 (66.7%, 4/6)were G145R mutation. Ratio of genotype C in occult donors (10/17) was statistically higher than HBs-positive donors (0/15, P<0.01), which implied that HBV genotype C leaded to occult infection more easily.

Quantitation of HTLV-I proviral load using real-time quantitative PCR with Taqman MGB probe / 病毒学报

A quantitative real-time PCR assay was developed to measure the proviral load of human T-lymphotropic virus type I (HTLV-I) in peripheral blood. The technology utilizes special primers and Taqman MGB fluorescence probe to measure amplification products from the gag-pro-pol polyprotein gene of HTLV-I. HTLV-I copy number was normalized to the amount of cellular DNA by quantitation of the beta-actin gene, The amplification system was sensitive to detect 5 copy/microL. The standard curve had a good linearity when the quantity for the gene was between 10(3) and 10(7) copy/microL (R2 = 0.999). Good reproducibility was observed in each intra- and inter-assay. We also measured proviral load in peripheral blood in 12 HTLV-I seropositive former blood donors. Proviral load for HTLV-I infected donors ranged from 0.015 to 12.819 copy/cell in WBC with the mean of 3.116 copy/cell.

TNF-alpha induced reversal of multidrug resistance in human hepatocellular carcinoma cells / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the reversal of multidrug resistance in the cell line HepG2/ADM induced by TNF-alpha.</p><p><b>METHODS</b>HepG2/ADM cells were incubated with different concentrations of TNF-alpha (100, 500 and 2500 U/ml) for 72 h. Real-time PCR was performed to compare the mRNA levels of MDR1 with PPAR-alpha in the different concentrations of TNF-alpha treated cells. The Annexin V assay was used to check cell apoptosis induced by 0.5 mg/L adriamycin. Rhodamine 123 efflux assay and MTT assay were used to study P-gp activity and drug resistance in each group, respectively.</p><p><b>RESULTS</b>TNF-alpha could induce down-regulation of MDR1 and up-regulation of PPAR-alpha. Meanwhile, it could enhance cell cytotoxicity and cell apoptosis induced by 0.5 mg/L adriamycin.</p><p><b>CONCLUSIONS</b>TNF-alpha could partially reverse the multidrug resistance of HepG2/ADM cells by down-regulating the expression of MDR1 and up-regulating the expression of PPAR-alpha.</p>

Hydrophobicity of reactive site loop of SCCA1 affects its binding to HBV / 生物工程学报

Squamous cell carcinoma antigen 1 (SCCA1), a member of the ovalbumin family of serine protease inhibitors, includes several variants. It was reported that expression of two SCCA1 (BP and AJ515706) in cells results in increased binding of HBV to these cells by the interaction of the expressed BP and AJ515706 with HBV pre-S1 domain. In this study, a SCCA1 (A1) was isolated from HepG2, but it appears to lack this ability. A possible role of two mutants, A1-BP and BP-A1, constructed by interchanging the carboxyl terminal of A1 and BP, was investigated. Cells expressing A1-BP rather than BP-A1 showed an increased virus binding capacity. Comparison of A1 sequence with the sequence of BP indicated the presence of only three amino acid changes in the carboxyl terminal, two of them in the reactive site loop (RSL) of SCCA1. Primary structure analysis revealed that the hydrophobicity of BP and AJ515706 in this domain is higher than that of A1. Changing the aa349 of A1 from low hydrophobic glutamic acid to high hydrophobic valine enhanced HBV binding. In contrast, changing the aa349 of BP from valine to glutamic acid reduced HBV binding. Our finding suggests that the hydrophobicity of RSL of SCCA1 may play an important role in HBV binding to cells.

Detection of hepatitis E virus on a blood donor and its infectivity to rhesus monkey / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To understand the infectivity and pathogenicity of the plasma of hepatitis E virus (HEV) viremia to primate animals.</p><p><b>METHODS</b>RNA fragment of HEV genotype IV was detected on one healthy donor who was positive for anti-HEV IgM and negative for anti-HEV IgG. Then 10 ml plasma from above donor was transfused to rhesus monkey to observe its infectivity and pathogenicity.</p><p><b>RESULTS</b>Acute hepatitis E was developed in rhesus monkey who accept HEV RNA positive plasma. It was confirmed by virological, immunological, biochemical and histopathological data.</p><p><b>CONCLUSION</b>Acute hepatitis E can be induced by plasma transfusion of HEV viremia, which indicate the possibility of transfusion transmitted hepatitis E</p>