Asymmetric figure-of-eight single-layer suture technique for intestinal anastomosis: A preliminary study.

Background: Anastomotic leakage is a life-threatening complication. Improvement of the anastomosis technique is needed, especially in patients with an inflamed edematous intestine. The aim of our study was to evaluate the safety and efficacy of an asymmetric figure-of-eight single-layer suture technique for intestinal anastomosis in pediatric patients. Methods: A total of 23 patients underwent intestinal anastomosis at the Department of Pediatric Surgery of Binzhou Medical University Hospital. Demographic characteristics, laboratory parameters, anastomosis time, duration of nasogastric tube placement, day of first postoperative bowel movement, complications, and length of hospital stay were statistically analyzed. The follow-up was conducted for 3-6 months after discharge. Results: Patients were divided into two groups: the single-layer asymmetric figure-of-eight suture technique (group 1) and the traditional suture technique (group 2). Body mass index in group 1 was lower than in group 2 (14.43 ± 3.23 vs. 19.38 ± 6.74; P = 0.036). The mean intestine anastomosis time in group 1 (18.83 ± 0.83 min) was less than that in group 2 (22.70 ± 4.11 min; P = 0.005). Patients in group 1 had an earlier first postoperative bowel movement (2.17 ± 0.72 vs. 2.80 ± 0.42; P = 0.023). The duration of nasogastric tube placement in group 1 was shorter than that in group 2 (4.12 ± 1.42 vs. 5.60 ± 1.57; P = 0.043). There was no significant difference in laboratory variables, complication occurrence, and length of hospital stay between the two groups. Conclusion: The asymmetric figure-of-eight single-layer suture technique for intestinal anastomosis was feasible and effective. More studies are needed to compare the novel technique with the traditional single-layer suture.

Clinical Application and Synthesis Methods of Deuterated Drugs.

Many drugs have adverse absorption, distribution, metabolism, and excretory (ADME) properties that prevent their widespread use or limit their use in some indications. In addition to preparation techniques and prodrug strategies, deuterium modification is a viable method for improving ADME properties. Deuterated drugs have attracted increasing attention from the pharmaceutical industry in recent years. To date, two deuterated drugs have been approved by the FDA. In 2017, austedo was approved by the FDA as a new drug for Huntington's disease in the United States, the first deuterium drug to be marketed worldwide. Recently (June 9, 2021), donafinil has been listed in China; this result has caused major pharmaceutical companies and the pharmaceutical industry to pay attention to deuterium technology again. In addition, BMS-986165, RT001, ALK-001, HC-1119, AVP-786 and other drugs are in phase III clinical studies, and some solid deuterium compounds have entered phase I and II clinical trials. The deuterium strategy has been widely used in pharmaceutical research and has become a hot spot in pharmaceutical research in recent years. In this paper, the research and development of deuterated drugs are reviewed, and the influence of deuterium modification on drugs, the advantages of deuterium strategies and the synthesis strategies of deuterated drugs are mainly introduced. Hoping to provide references for clinical application, the discovery of new deuterium chemical entities and research and development of new deuterated drugs.

The use of carcinoembryonic antigen levels to predict lung nodule malignancy: a meta-analysis.

OBJECTIVES: To assess the diagnostic value of serum carcinoembryonic antigen (CEA) as a diagnostic biomarker that can be used to differentiate between benign and malignant lung nodules (LNs). METHODS: PubMed, Cochrane Library, and Embase were reviewed from January 2000 to April 2020 for eligible studies. Stata v12.0 was used to conduct this meta-analysis. RESULTS: Our initial literature search identified 511 potentially relevant studies, of which 11 were ultimately included in the present meta-analysis. Ten studies were retrospective and only 1 study was prospective. Overall these studies incorporated 2760 patients and 2760 total LNs (1733 malignant, 1027 benign). Pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), and diagnostic odds ratio (DOR) values for these studies were 0.33 (95% CI: 0.20-0.49), 0.92 (95% CI: 0.85-0.96), 3.96 (95% CI: 2.84-5.54), 0.73 (95% CI: 0.62-0.87), and 5.42 (95% CI: 3.77-7.78), respectively. The area under curve (AUC) value was 0.77, consistent with moderate diagnostic accuracy. We detected significant heterogeneity when calculating pooled sensitivity (I2 = 95.9%, P = 0.00), specificity (I2 = 92.0%, P = 0.00), PLR (I2 = 61.7%, P = 0.00), NLR (I2 = 92.8%, P = 0.00), and DOR (I2 = 93.8%, P = 0.00). No significant evidence of publication bias was detected via Deeks' funnel plot asymmetry test (P = 0.371). Meta-regression analysis revealed different reference standards to be closely associated with both sensitivity and specificity. CONCLUSIONS: Serum CEA can achieve moderate diagnostic performance as a means of differentiating between malignant and benign LNs.

Research Status, Synthesis and Clinical Application of Recently Marketed and Clinical BCR-ABL Inhibitors.

Tyrosine kinases expressed by BCR-ABL fusion genes can cause changes in cell proliferation, adhesion, and survival properties, which are the main causes of chronic myelogenous leukemia (CML). Inhibiting the activity of BCR-ABL tyrosine kinase has become one of the effective methods for the treatment of chronic myelogenous leukemia. Initially, imatinib was the first small molecule of BCR-ABL tyrosine kinases inhibitors (TKIs) for the effective treatment of chronic myelogenous leukemia. Later, due to the emergence of various BCR-ABL mutations, especially T315I mutation, imatinib developed strong resistance. The second-generation kinase inhibitors dasatinib and nilotinib were able to overcome most of the mutation resistance but not T315I mutations. Therefore, in order to further overcome the problem of drug resistance, new types of KTIs such as flumatinib and radotinib have been developed, providing more options for clinical treatment. Some new drugs have entered clinical trials. In this review, two new BCRABL inhibitors (flumatinib and radotinib) and five new BCR-ABL inhibitors have been introduced into the clinical market in recent years. We reviewed their research status, synthesis methods, and clinical applications.

SPP1 and FN1 are significant gene biomarkers of tongue squamous cell carcinoma.

Tongue squamous cell carcinoma (TSCC) is one of the most common malignant tumor types in the oral and maxillofacial region. The etiology and pathogenesis behind TSCC is complicated. In the present study, three gene expression profiles, namely GSE31056, GSE13601 and GSE78060, were downloaded from the Gene Expression Omnibus (GEO). The GEO2R online tool was utilized to identify differentially expressed genes (DEGs) between TSCC and normal tissue samples. Furthermore, a protein-protein interaction (PPI) network was constructed and hub genes were validated and analyzed. A total of 83 common DEGs were obtained in three datasets, including 48 upregulated and 35 downregulated genes. Pathway enrichment analysis indicated that DEGs were primarily enriched in cell adhesion, extracellular matrix (ECM) organization, and proteolysis. A total of 63 nodes and 218 edges were included in the PPI network. The top 11 candidate hub genes were acquired, namely plasminogen activator urokinase (PLAU), signal transducer and activator of transcription 1, C-X-C motif chemokine ligand 12, matrix metallopeptidase (MMP) 13, secreted phosphoprotein 1 (SPP1), periostin, MMP1, MMP3, fibronectin 1 (FN1), serpin family E member 1 and snail family transcriptional repressor 2. Overall, 83 DEGs and 11 hub genes were screened from TSCC and normal individuals using bioinformatics and microarray technology. These genes may be used as diagnostic and therapeutic biomarkers for TSCC. In addition, SPP1 and FNl were identified as potential biomarkers for the progression of TSCC.

Benign pathologies results from lung nodule percutaneous biopsies: How to differentiate true and false benign?

OBJECTIVES: The objective was to identify predictors of true negatives in lung nodules (LNs) with computed tomography-guided percutaneous biopsy (CTPB)-based benign pathological results. MATERIALS AND METHODS: We included 90 total patients between January 2013 and December 2017 that had CTPB-based nonspecific benign pathologies and used these patients as a training group to accurately identify true-negative predictors. A validation group of 50 patients from January 2018 to June 2019 to confirm predictor reliability. RESULTS: CTPB was conducted on 90 LNs from the training group. True-negative and false-negative CTPB-based pathologies were obtained for 79 and 11 LNs, respectively. CTPB-based benign results had a negative predictive value of 87.8% (79/90). Univariate and multivariate analyses revealed younger age (P = 0.019) and CTPB-based chronic inflammation with fibroplasia (P = 0.010) to be true-negative predictors. A predictive model was made by combining these two prognostic values as follows: score = -7.975 + 0.112 × age -2.883 × CTPB-based chronic inflammation with fibroplasia (0: no present; 1: present). The area under receiver operator characteristic (ROC) curve was 0.854 (P < 0.001). To maximize sensitivity and specificity, we selected a cutoff risk score of -0.1759. The application of this model to the validation group yielded an area under the ROC curve of 0.912 (P < 0.001). CONCLUSIONS: Our predictive model showed good predictive ability for identifying true negatives among CTPB-based benign pathological results.

Measurement and prediction on thermal conductivity of fused quartz.

Thermal conductivity of soil is a basic physical property related to heat conduction, and also is one of parameters widely applied in geotechnical engineering. The effect of gradation on the thermal conductivity of fused quartz was analyzed by thermal needle tests. The different particle size with the same uniformity coefficient (Cu = 3.2) and different uniformity coefficient for the same particle size (0.10~1.00 mm) were considered in this study. It shows that the thermal conductivity of fused quartz decreases with the decreasing of the mean particle size and with the increasing of the porosity. Simple modified methods to estimate the value of thermal conductivity are proposed, and had been demonstrated successfully by conducting fused quartz, carbonate sand and Ottawa sand.

Alternative technique to save ischemic bowel segment in management of neonatal short bowel syndrome: A case report.

BACKGROUND: Congenital short bowel syndrome (SBS) associated with malrotation, gut volvulus and jejuno-ileal atresia is a very rare condition. It is a severe challenge for surgeons to preserve residual ischemic bowel segment in the management of short bowel syndrome,especially in neonates. CASE SUMMARY: We report a newborn baby with gut malrotation associated with jejuno-ileal atresia, congenital SBS and jejunal volvulus. Hematemesis and abdominal distention were noted. At laparotomy, malrotation associated with jejuno-ileal atresia, congenital SBS and jenunal volvulus was confirmed. The total length of the small bowel was 63 cm with proximal jejunal bowel segment measuring 38 cm, including 18 cm necrotic segment below the Treitz's ligament and 20 cm severe ischemic segment. The distal part of the small bowel was 25 cm in length and only about 0.8 cm in diameter. Ladd's procedure, necrotic segment resection and end-to-back duodeno-ileal anastomosis were performed. The residual severe ischemic jejunum was preserved with single proximal stoma and distal end closure. Three months later, to restore the continuity of the isolated gut segment, end-to-end duodeno-jejunal and jejuno-ileal anastomosis was performed. The entire functional small bowel length increased to 80 cm. Intravenous fluid therapy and parenteral nutrition were discontinued on the 10th day postoperatively. Twelve months later, her body weight was 9.5 kg. CONCLUSION: Isolation of severe ischemic bowel segment and staged anastomosis to restore the gut continuity for infants with SBS are safe and feasible.

Au@SiO2@CuInS2-ZnS/Anti-AFP fluorescent probe improves HCC cell labeling.

BACKGROUND: Clear tumor imaging is essential to the resection of hepatocellular carcinoma (HCC). This study aimed to create a novel biological probe to improve the HCC imaging. METHODS: Au nano-flower particles and CuInS2-ZnS core-shell quantum dots were synthesized by hydrothermal method. Au was coated with porous SiO2 and combined with anti-AFP antibody. HCC cell line HepG2 was used to evaluate the targeting efficacy of the probe, while flow cytometry and MTT assay were used to detect the cytotoxicity and bio-compatibility of the probe. Probes were subcutaneously injected to nude mice to explore light intensity and tissue penetration. RESULTS: The fluorescence stability of the probe was maintained 100% for 24 h, and the brightness value was 4 times stronger than that of the corresponding CuInS2-ZnS quantum dot. In the targeting experiment, the labeled HepG2 emitted yellow fluorescence. In the cytotoxicity experiments, MTT and flow cytometry results showed that the bio-compatibility of the probe was fine, the inhibition rate of HepG2 cell with 60% Cu-QDs/Anti-AFP probe and Au-QDs/Anti-AFP probe solution for 48 h were significantly different (86.3%±7.0% vs. 4.9%±1.3%, t = 19.745, P<0.05), and the apoptosis rates were 83.3%±5.1% vs. 4.4%±0.8% (P<0.001). In the animal experiment, the luminescence of the novel probe can penetrate the abdominal tissues of a mouse, stronger than that of CuInS2-ZnS quantum dot. CONCLUSIONS: The Au@SiO2@CuInS2-ZnS/Anti-AFP probe can targetedly recognize and label HepG2 cells with good bio-compatibility and no toxicity, and the strong tissue penetrability of luminescence may be helpful to surgeons.

The associations between MDM4 gene polymorphisms and cancer risk.

Considerable studies have investigated the associations between MDM4 gene polymorphisms and cancer risk recently, but with contradictory results. The aim of this meta-analysis was to evaluate the associations between MDM4 gene polymorphisms and cancer risk. Relevant studies were identified by a systematic search of PubMed, Embase, and CNKI databases. Crude odds ratios (ORs) and 95% confidence intervals (CIs) were used to describe the strength of the associations. Fifty-six studies published in 11 publications involving 18,910 cases and 51,609 controls were included in this meta-analysis. Five MDM4 gene polymorphisms were evaluated: rs4245739, rs1563828, rs11801299, rs10900598, and rs1380576. Our analyses suggested that the rs4245739 polymorphism was significantly associated with overall cancer risk. Furthermore, stratification analyses of ethnicity indicated that rs4245739 decreased the risk of cancer among the Asian population, and stratification analyses of smoking status indicated that rs4245739 decreased the risk of cancer among nonsmokers. However, stratification analyses of cancer type and sex suggested that rs4245739 was not related to cancer risk. There were no associations of rs1563828, rs11801299, rs10900598, or rs1380576 with overall cancer risk. In conclusion, our analyses indicated that rs4245739 polymorphism in the MDM4 gene may play an important role in the etiology of cancer.

MYO9B gene polymorphisms are associated with the risk of inflammatory bowel diseases.

Myosin IXB (MYO9B) gene polymorphisms have been extensively investigated in terms of their associations with inflammatory bowel disease (IBD), with contradictory results. The aim of this meta-analysis was to evaluate associations between MY09B gene polymorphisms and the risk of IBD, Crohn's disease (CD) and ulcerative colitis (UC). Eligible studies from PubMed, Embase, and CNKI databases were identified. Pooled odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. Ten studies published in eight papers reporting 8,975 cases and 9,482 controls were included in this meta-analysis. Five MY09B gene polymorphisms were evaluated: rs1545620, rs962917, rs1457092, rs2305764, and rs2305767. Our data suggested that the rs1545620 polymorphism was associated with a decreased risk of IBD. A similar result was found for rs2305767 and UC. The rs962917 single nucleotide polymorphism (SNP) increased the risk of IBD, CD and UC. Moreover, rs1457092 increased the risk of IBD and UC. Rs2305764 was also associated with an increased risk of IBD. Furthermore, stratification analyses indicated that rs1545620 decreased the risk of IBD, while rs962917 increased the risk of IBD, CD and UC in Caucasian populations. To sum up, our data indicate that these five SNPs in MY09B are significantly associated with the risk of IBD.

Bio-compatibility and cytotoxicity studies of water-soluble CuInS2-ZnS-AFP fluorescence probe in liver cancer cells.

BACKGROUND: The oncogenesis of hepatocellular carcinoma (HCC) is not clear. The current methods of the pertinent studies are not precise and sensitive. The present study was to use liver cancer cell line to explore the bio-compatibility and cytotoxicity of ternary quantum dots (QDs) probe and to evaluate the possible application of QDs in HCC. METHODS: CuInS2-ZnS-AFP fluorescence probe was designed and synthesized to label the liver cancer cell HepG2. The cytotoxicity of CuInS2-ZnS-AFP probe was evaluated by MTT experiments and flow cytometry. RESULTS: The labeling experiments indicated that CuInS2-ZnS QDs conjugated with AFP antibody could enter HepG2 cells effectively and emit intensive yellow fluorescence by ultraviolet excitation without changing cellular morphology. Toxicity tests suggested that the cytotoxicity of CuInS2-ZnS-AFP probe was significantly lower than that of CdTe-ZnS-AFP probe (t test, F=0.8, T=-69.326, P<0.001). For CuInS2-ZnS-AFP probe, time-effect relationship was presented in intermediate concentration (>20%) groups (P<0.05) and dose-effect relationship was presented in almost all of the groups (P<0.05). CONCLUSION: CuInS2-ZnS-AFP QDs probe had better bio-compatibility and lower cytotoxicity compared with CdTe-ZnS-AFP probe, and could be used for imaging the living cells in vitro.

Association of IL12B Gene Polymorphisms with Rheumatoid Arthritis: A Meta-analysis.

BACKGROUND AND AIMS: Currently published papers regarding the relationship between interleukin (IL)-12B gene polymorphisms and rheumatoid arthritis (RA) are contradictory. The aim of this meta-analysis was to evaluate the associations between the IL-12B gene polymorphisms (rs3122227 and rs6887695) and RA risk. METHODS: We searched PubMed, Embase, the Cochrane Library and the China Knowledge Resource Integrated Database. Pooled odds ratios (ORs) and 95% confidence intervals (95% CIs) were used to assess associations between IL12B gene polymorphisms and RA. RESULTS: A total of eight publications (4,409 cases and 5,591 controls) were included in this meta-analysis. The results demonstrated that rs3122227 and rs6887695 were not associated with RA risk based on current included studies. However, stratification analyses indicated rs6887695 was associated with RA in Asian patients. Rs3122227 was not related with RA in Asian or Caucasian patients. CONCLUSIONS: Our data indicated that IL-12B gene polymorphisms were not related with RA. However, rs6887695 was associated with RA in Asian patients. Further larger-scale studies are urgently needed to identify the association between IL-12B gene polymorphisms and RA in Asian populations.

Synthesis and antitumor activity of novel 2-substituted indoline imidazolium salt derivatives.

A series of novel 2-substituted indoline imidazolium salt derivatives has been prepared and evaluated in vitro against a panel of human tumor cell lines. The results suggest that the existence of a substituted benzimidazole ring and substitution of the imidazolyl-3-position with a naphthylacyl or 2-naphthylmethyl group were vital for modulating the cytotoxic activity. Compound 25 was found to be the most potent derivative with IC50 values of 0.24-1.18 µM, and exhibited cytotoxic activity selectively against MCF-7, SW480, SMMC-7721 and HL-60 cell lines, while compound 26 showed powerful inhibitory activities selectively against SMMC-7721 and A549 cell lines. Compound 25 can induce G2/M phase cell cycle arrest and apoptosis in SMMC-7721 cells.

Synthesis and cytotoxic activity of novel 1-((indol-3-yl)methyl)-1H-imidazolium salts.

A series of novel 1-((indol-3-yl)methyl)-1H-imidazolium salts were prepared and evaluated in vitro against a panel of human tumor cell lines. The results suggest that the 5,6-dimethyl-benzimidazole ring, and substitution of the imidazolyl-3-position with a naphthylacyl or 4-bromophenacyl group, were vital for modulating inhibitory activity of cell growth. In particular, 1-((N-Boc-indol-3-yl)methyl)-3-(2-naphthylacyl)-1H-5,6-dimethyl-benzimidazolium bromide was found to be the most potent derivative and more selective against myeloid liver carcinoma (SMMC-7721), lung carcinoma (A549) and breast carcinoma (MCF-7), with IC50 values 1.9-fold, 1.7-fold and 4.8-fold lower than DDP. This compound can induce significant cell apoptosis in SMMC-7721 cells.

[Analysis of aerosol concentration characteristics under continuous synoptic systems in Qingdao].

Based on weather data and data obtained by Particle Sizer GRIMM180 set up in Qingdao, aerosol was qualitively classified into sea-fog aerosol, refreshing aerosol and suspended dust aerosol. Analysis of mass concentration and number concentration of three different kinds of aerosols was conducted, and the results are shown as below: 1) total mass concentration of different kinds of aerosol is obviously different; 2) sea-fog aerosol primarily includes particles of which size ranging from 1 microm to 2.5 microm, and refreshing aerosol, includes particles of which size less than 1 microm, and suspended dust aerosol, includes particles of which size ranging from 2.5 microm to 10 microm. 3) precipitation has important role on decreasing larger particle concentration and increasing tiny particle concentration. 4) those tiny particles of which size is less than 1 microm, especially less than 0.6 microm, show an activation phenomenon when they located before the surface weather systems, where air humidity is considerable high. 5) trend of number concentration variability of different particles has different characteristic modes when aerosol property has been changed.

Activated changes of platelet ultra microstructure and plasma granule membrane protein 140 in patients with non-small cell lung cancer.

BACKGROUND: Non-small cell lung cancer (NSCLC) is the leading cause of cancer mortality worldwide. Platelet activation may play an important role in pathologic progress in lung cancer. In this study, we aimed to clarify the influence of activated platelets on lung cancer generation and growth, and the relationship among these functional and ultrastructural changes of platelets and the severity of pathogenetic condition in these patients with NSCLC. METHODS: One hundred and thirty-six cases of patients with pathologically confirmed NSCLC were included in this study. Fifty-four healthy people were enrolled as controls. The change of ultra microstructure and activity of blood platelets were observed under the transmission and scanning electron microscope. Simultaneous determination of plasma granule membrane protein 140 (GMP-140) was made. RESULTS: Transmission electron microscopy showed remarkable changes of ultra microstructure of platelets in patients with NSCLC, including swelling, increase of a-granules, vesicles, and glycogenosome. Scanning electron microscopy showed many more surface processes and wrinkles on platelets in patients with NSCLC. The reference plasma levels of GMP-140 of healthy controls were (18.2 +/- 2.7) microg/L. The plasma levels of GMP-140 in patients with NSCLC were (47.8 +/- 12.3) microg/L, which were much higher than those of the controls. There was a medium positive correlation between plasma levels of GMP-140 and amount of a-granules (r = 0.514, P < 0.01) and a high positive correlation between plasma levels of GMP-140 and area of platelet (r = 0.84, P < 0.01) in patients with NSCLC. The Kaplan-Meier survival curve analysis showed significant shift to the left in patients with NSCLC whose a-granules per platelet were 19 or more compared to those 18 or less (Log rank statistic, chi(2) = 17.38, P < 0.01). CONCLUSIONS: There are significant activated changes of ultra microstructure and increased activity of blood platelets in patients with NSCLC. These activated platelets may play an important role in the generation and growth of lung cancer. These changes can be used as a diagnostic index of severity, progression, and prognosis of NSCLC.

Raman analysis of CdSe/CdS core-shell quantum dots with different CdS shell thickness.

We report the first-order and high-order Raman scattering from core-shell CdSe/CdS nanocrystals and investigate the evolution of the longitudinal mode of CdSe (LO1) and CdS (LO2), and the surface mode of CdSe (SO1) and CdS (SO2) with increasing shell thickness (0-5.5 ML (monolayer)). We find that the shift of the LO2 peak from 268 to 291 cm(-1) agrees well with the theoretical values based on the phonon confinement model. The variation of the dielectric environment of the CdSe core with increasing CdS shell thickness is modified according to the shift of SO1 from 198 to 185 cm(-1). The SO2 modes at 267 and 275 cm(-1), corresponding to the shell thickness 3.5 and 5.5 ML in CdSe/CdS nanocrystals, respectively, are also obtained for the first time in our experiment. Moreover, they agree well with the theoretical values of the dielectric corresponding function model. Besides, a new Raman peak at 482 cm(-1) is observed, and it remains at that value with shell growth, the new peak is supposed to be caused by the alloying at the core-shell interface. Therefore, Raman spectroscopy can be used to determine the shell thickness and other surface and interface parameters of CdSe/CdS core-shell nanocrystals.

[Expression and identification of HCV core protein in human hepatocytes].

AIM: To construct the recombinant plasmid of HCV core protein, and to express and identify it in normal human hepatocyte HL-7702. METHODS: HCV core gene was cloned by using PCR from plasmid pBRTM/HCV1-3011 which included the full length of HCV gene. The core segment with expression plasmid pcDNA3.1(-) was recombined to construct eukaryotic expression plasmid pcDNA3.1(-)/core, which was then transfected into human hepatocytes by using poly-cation. The expression of core protein was detected by immunochemical staining and Western blot. RESULTS: The length and sequence of the cloned core segment were correct. The transfected HL-7702 cells expressed the core protein. CONCLUSION: The eukaryotic expression plasmid pcDNA3.1(-)/core including HCV core gene is successfully constructed. The effective expression of HCV core protein in human hepatocytes is useful for further development of HCV core antigen.

Construction of eukaryotic expression plasmid containing HCV NS3 segment and protein expression in human HL-7702 hepatocytes.

AIM: To construct the eukaryotic expression plasmid containing HCV NS3 segment and to analyze the expression of NS3 protein in normal human hepatocyte HL-7702. METHODS: We amplified HCV NS3 fragment from plasmid pBRTM/HCV 1-3011 containing the whole length of HCV genome, recombined it with expression vector pcDNA3.1(-) to form the eukaryotic expression vector pcDNA3.1(-)/NS3, and transfected human HL-7702 hepatocytes with the recombined plasmid by cationic polymers. The expressed HCV NS3 protein was detected and analyzed by immunohistochemical method and Western blot. RESULTS: The amplified NS3 fragments had correct molecule weight and sequence. The successfully constructed eukaryotic expression plasmids were transfected to HL-7702 cells. The expressed NS3 proteins had correct molecular weight 70000. CONCLUSION: Eukaryotic expression vector pcDNA3.1 (-)/NS3 containing NS3 segment of HCV can be constructed, the sequence of NS3 fragments is consistent with the template. Normal human HL-7702 hepatocytes can efficiently express specific HCV NS3 protein in vitro.