Background and purpose:The reason for hepatitis B virus (HBV) with hepatocyte specificity is PreS1 enchased on the hepatitis B virus envelope (HBVE). So HBVE may have a potential application in liver targeting gene transfer. In this study, we investigated whether HBVE has the ability to target liver cancer cells. Methods:HBVE was obtained from the supernatant of Hep G 2.2.15 cells through PEG8000 system and ?-propiolactone method. The pIRS2-EGFP was packed with HBVE and resulted in the product HBVE-GFP. HBVE-GFP was transfected into HepG2, A549, HeLa and FB cells. The green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results:The GFP could be observed in the four groups, but the HepG2 group had a higher fluorescent intensity than the other 3 groups. The transfected rate of HepG2 group was (71.35?0.03)% , much higher than other groups(P

Objective: To construct a liver targeting gene transfer vector using hepatitis B virus envelope particles. Methods: Hepatitis B viruses were obtained from the supernatant of HepG 2.2.15 cells by a PEG8000 system and were inactivated by ?-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES_2-EGFP to assess their packing ability. Subsequently, the products were studied with ELISA, PCR, SDS-PAGE, and electron microscopy. Finally, the product was used to transfect HepG2 cells and the green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results: The acquired hepatitis B virus envelope retained the surface protein HBsAg+pre S_1+pre S_2, but with no virus DNA. The prepared envelope had high packing ability for GFP and the packed GFP had a high transfection rate in HepG2 cell. Conclusion: Hepatitis B virus envelope has been successfully obtained from the supernatant of HepG 2.2.15 cells with a PEG8000 system and ?-propiolactone.

Objective:To observe the transfection efficacy and targeting efficiency of hepatitis B virus envelope(HBVE) as a gene transfer vector for liver cancer cells.Methods:HBVE was obtained from the supernatant of HepG2.2.15 cells with a PEG8000 system and ?-propiolactone.The pIRS2-EGFP was packed with HBVE to obtain HBVE-GFP and was packed with liposome to obtain Liposome-GFP.HBVE-GFP and Liposome-GFP were used to transfect human hepatoblastoma cell line HepG 2 to study the transfection efficiency.HepG 2,A 549,HeLa and FB cells were transfected with HBVE-GFP to appraise the targeting ability of HBVE-GFP.GFP protein expression was observed under a fluorescent microscope and the ratio of GFP positive cells was determined by flow cytometry.Results:(1) Transfection efficiency:The GFP protein was observed in both the liposome group and the HBVE group under the fluorescent microscope;the fluorescent intensity in the HBVE group was 3-4 times that of liposome group as determined by flow cytometry(P

BACKGROUND: In the treatment of osteoporotic vertebral compression fractures with percutaneous vertebroplasty, the efficacy and safety of bone cement injection by unipedicular and bipedicular approaches are still controversial. Some studies suggest that bone cement injection via unipedicular approach can shorten operation time and reduce postoperative complications, while the other studies suggest that bone cement injection via bipedicular approach can make bone cement distribute more evenly in the vertebral body and relieve pain better. OBJECTIVE: To systematically assess the efficacy and safety of percutaneous vertebroplasty via unipedicular versus bipedicular approach in the treatment of osteoporotic vertebral compression fractures. METHODS: Randomized controlled trials about unipedicular versus bipedicular percutaneous vertebroplasty in the treatment of osteoporotic vertebral compression fracture published before September 18 t h, 2018 were retrieved in the PubMed, Cochrane library, Embase, CNKI, VIP, WanFang data and CBM. Two researchers independently screened all the literatures, carried out data extraction and used improved Jadad to evaluate the methodological quality of the included studies. Meta-analysis using Revam 5.3 was conducted. Egger's test was utilized to evaluate the publication bias. RESULTS AND CONCLUSION: Totally 14 randomized controlled trials including 900 cases were eventually included, 452 cases in unipedicular approach group and 448 cases in bipedicular approach group. The Meta-analysis results showed that compared with the bipedicular approach, the unipedicular approach required shorter operation time [weighted mean difference (WMD) =-16.59, 95％ confidence interval (CI) (-19.25, -13.94), P < 0.001], smaller amount of bone cement injected [WMD=-1.27, 95％ CI (-1.64, -0.89), P < 0.001], and had lower incidence of cement leakage [relative risk=0.70, 95％ CI (0.53, 0.92), P =0.01]. There were no significant differences in short-and long-term Visual Analogue Scale scores, short-and long-term Oswestry Disability Index scores, and the postoperative incidence of adjacent vertebral fractures between the two groups (P> 0.05). Overall, bone cement injection both via bipedicular and unipedicular approaches can lead to a significant improvement in pain relief and living quality of osteoporotic vertebral compression fracture patients, but bone cement injection via unipedicular approach can shorten operation time, reduce cement volume and lower the incidence of cement leakage compared with the bipedicular approach.

Objective: To identify potential variant in a child diagnosed as infantile neuroaxonal dystrophy. Methods: Genomic DNA was extracted from peripheral blood samples from the patient and his parents and subjected to next generation sequencing. Suspected variant was verified by PCR and Sanger sequencing. Pathogenicity of the mutation was predicted by using bioinformatic software including SIFT and PolyPhen-2. Results: The child was found to carry compound heterozygous variations c. 668C>A (p.Pro223Gln) and c. 2266C>T (p.Gln756Ter) of the PLA2G6 gene, which were respectively inherited from his father and mother. c. 2266C>T has changed codon 756 (glutamine) into a stop codon, resulting premature termination of peptide chain synthesis. c. 2266C>T has not been reported previously and was predicted to be harmful. Conclusion The compound variants of c. 668C>A (p.Pro223Gln) and c. 2266C>T (p.Gln756Ter) of the PLA2G6 gene probably underlies the disease in the child. Above finding has enriched the variant spectrum of the PLA2G6 gene.

Objective:To explore the treatment effect of gefitinib on epidermal growth factor receptor (EGFR)-positive advanced non-small cell lung cancer (NSCLC).Methods:Sixty patients with EGFR-positive advanced NSCLC who were admitted to the 904th Hospital of Joint Logistics Support Force of Chinese PLA from March 2016 to January 2020 were selected. They were divided into gefitinib treatment group (30 cases, treated with gefitinib) and combined treatment group (30 cases, treated with pemetrexed combined with cisplatin) by random number table. The anti-tumor efficacy, levels of tumor markers [serum carcinoembryonic antigen (CEA), cytokeratin 19 fragment antigen (CYFRA21-1) and neuron-specific enolase (NSE)] before and after treatment, adverse reactions and 6-month overall survival (OS) rate were compared between the two groups.Results:The clinical control rate of gefitinib treatment group was higher than that of combined treatment group [76.7% (23/30) vs. 50.0% (15/30), χ2 = 4.593, P = 0.032]. There was no significant difference in the levels of CEA, CYFRA21-1 and NSE between the two groups before treatment (all P > 0.05). The levels of CEA, CYFRA21-1 and NSE after treatment in gefitinib treatment group were (902±41) μg/L, (3.1±0.4) ng/ml and (17.7±2.3) ng/ml. The levels of CEA, CYFRA21-1 and NSE after treatment in combined treatment group were (999±51) μg/L, (4.0±0.5) ng/ml and (19.4±3.1) ng/ml. The levels of CEA, CYFRA21-1 and NSE after treatment in gefitinib treatment group were lower than those in combined treatment group ( t = 7.441, P < 0.01; t = 7.459, P < 0.01; t = 2.486, P = 0.016).The levels of CEA, CYFRA21-1 and NSE after treatment in the two groups were all lower than those before treatment, and the differences were statistically significant (all P < 0.05). There was no significant difference in the incidence of rash, thrombocytopenia, digestive tract reaction, and proteinuria between the two groups [26.7% (8/30) vs. 23.3% (7/30), χ2 = 0.089, P = 0.766; 16.7% (5/30) vs. 13.3% (4/30), χ2 = 0.131, P = 0.718); 30.0% (9/30) vs. 26.7% (8/30), χ2 = 0.082, P = 0.774; 10.0% (3/30) vs. 13.3% (4/30), χ2 = 0.162, P = 0.688]. After 6 months of treatment, the OS rate in gefitinib treatment group was 93.3%, and that in combined treatment group was 83.3%, and there was no statistical difference between the two groups ( χ2 = 1.456, χ2 = 0.228). Conclusion:Gefitinib treatment for EGFR-positive advanced NSCLC patients can enhance the anti-tumor efficacy, reduce the content of tumor markers, and has good safety.

Objective: To analyze variations of TYR and P genes among 14 patients with clinically diagnosed oculocutaneous albinism. Methods: Potential variations of the TYR and P genes were detected by Sanger sequencing. Novel variations were predicted with bioinformatics software including SIFT and PolyPhen-2. Results: No variation was found in the TYR gene, while 9 types of variations were found in the P gene among the 14 patients, which included c. 803-3C>G (7/26), c. 1327G>A (p.Val443Ile) (5/26), c. 632C>T (p.Pro211Leu) (4/26), c. 1832T>C (p.Leu611Pro) (3/26), c. 1349C>A (p.Thr450Lys) (2/26), c. 2363C>T (p.Ser788Leu) (2/26), c. 2228C>T (p.Pro743Leu) (1/26), c. 1525A>G(p.Thr509Ala) (1/26), and c. 1349C>T(p.Thr450Met) (1/26). Only 1 heterozygous variation was detected in 2 families. c. 2363C>T (p.Ser788Leu), c. 1832T>C (p.Leu611Pro) and c. 1525A>G (p.Thr509Ala) were not reported previously and predicted as "harmful" to the protein function. Conclusion The main type of ocular albinism is oculocutaneous albinism type Ⅱ in Liuzhou region, where the most common variations of the P gene were c. 803-3C>G and c. 1327G>A (p.Val443Ile). Above finding has enriched the variation spectrum of the P gene.

OBJECTIVE: To detect variants of IVD gene among 4 neonates with suspected isovalerate acidemia in order to provide a guidance for clinical treatment. METHODS: 111 986 newborns and 7461 hospitalized children with suspected metabolic disorders were screened for acyl carnitine by tandem mass spectrometry. Those showing a significant increase in serum isovaleryl carnitine (C5) were analyzed for urinary organic acid and variants of the IVD gene. RESULTS: Four cases of isovalerate acidemia were detected, which included 2 asymptomatic newborns (0.018‰, 2/111 986) and 2 children suspected for metabolic genetic diseases (0.268‰, 2/7461). The formers had no obvious clinical symptoms. Analysis of acyl carnitine has suggested a significant increase in C5, and urinary organic acid analysis has shown an increase in isovaleryl glycine and 3-hydroxyisovalerate. Laboratory tests of the two hospitalized children revealed high blood ammonia, hyperglycemia, decreased red blood cells, white blood cells, platelets and metabolic acidosis. The main clinical manifestations have included sweaty foot-like odor, feeding difficulty, confusion, drowsiness, and coma. Eight variants (5 types) were detected, which included c.158G>A (p.Arg53His), c.214G>A (p.Asp72Asn), c.548C>T (p.Ala183Val), c.757A>G (p.Thr253Ala) and 1208A>G (p.Tyr403Cys). Among these, c.548C>T and c.757A>G were unreported previously. None of the variants was detected by next generation sequencing of 2095 healthy newborns, and all variants were predicted to be likely pathogenic based on the guidelines from the American College of Medical Genetics and Genomics. CONCLUSION The incidence of isovalerate acidemia in Liuzhou area is quite high. Screening of metabolic genetic diseases is therefore recommended for newborns with abnormal metabolism. The discovery of novel variants has enriched the mutational spectrum of the IVD gene.
Infant, Newborn ; Child ; Humans ; Acidosis ; Carnitine ; Erythrocytes ; High-Throughput Nucleotide Sequencing