[Impact of nucleosides analogues and nucleotide analogues on the outcomes related to chronic hepatitis B based on non-antiviral effects].

Nucleoside analogues and nucleotide analogues can not only achieve long-term viral suppression in the treatment of most CHB patients but also have a positive impact on other CHB therapeutic goals and an improved prognosis. A certain difference can be observed in the impact of nucleotide analogues such as TDF and TAF and nucleoside analogues such as ETV on the clinical outcomes of CHB. Studies on the mechanism of action indicate that apart from inhibiting the direct antiviral effects of HBV reverse transcriptase, these two categories of drugs exhibit distinct impacts on immune-related signaling pathways, gene expression, genome stability, and other non-antiviral mechanisms. This article reviews the evidence on the potential non-antiviral mechanism of action of nucleoside analogues and nucleotide analogues and proposes a preliminary explanation for the observation trend of nucleotide analogues having a comparative advantage in clinical outcomes in CHB patients based on the latest research advancement.

[Epidemiological analysis and screening mode of breast cancer screening in Fujian province from 2015 to 2018].

Objective: To analyze the screening results of breast cancer in rural women in Fujian Province from 2015 to 2018, and to explore the screening mode of breast cancer. Methods: Breast cancer screening was provided for 35-64 years old rural women in Fujian province. Color Doppler ultrasound was used as the primary screening method. Suspected patients were referred to mammography. Color Doppler ultrasound and breast X -ray 4-5 grade were diagnosed by pathology. Results: The incidence of breast cancer from 2015 to 2018 was 56.96/10 million, 94.41/10 million, 71.61/10 million, and 73.05/10 million, respectively. And the peak age of breast cancer was (46-55) years. From 2015 to 2018, the proportion of cancer in situ showed an overall upward trend. The sensitivity of color Doppler ultrasound was 79.06%, and the sensitivity of mammography for those diagnosed abnormally by ultrasound was 91.88%. Conclusion: Breast cancer screening for women of the right age is an important measure to raise the rate of early diagnosis and early treatment and reduce the mortality of breast cancer. Screening mode based on breast ultrasound is the most suitable technology. We need to further expand the coverage of screening, improve the screening system and process, and ensure the health of women.

Genome-wide analysis of differentially expressed lncRNA in sporadic parathyroid tumors.

Diagnosis of parathyroid carcinoma on histological examination is challenging. Thousands of differentially expressed lncRNAs were identified on the microarray data between parathyroid cancer and adenoma samples. Four lncRNAs were significantly dysregulated in further validation. The "lncRNA score" calculated from these lncRNAs differentiated parathyroid carcinomas from adenomas. LncRNAs serve as biomarkers for parathyroid cancer diagnosis. INTRODUCTION: Diagnosis of parathyroid carcinoma (PC) on histological examination is challenging. LncRNA profile study was conducted to find diagnostic biomarkers for PC. METHODS: LncRNA arrays containing 91,007 lncRNAs as well as 29,857 mRNAs were used to assess parathyroid specimen (5 carcinomas and 6 adenomas). Bioinformatics analyses were also conducted to compare the microarray results between parathyroid carcinomas and adenomas (PAs). Differentially expressed lncRNAs of 11 PCs and 31 PAs were validated by real-time quantitative PCR. RESULTS: On the microarray data between PC and PA samples (fold change ≥ 2, P < 0.05), 1809 differentially expressed lncRNAs and 1349 mRNAs also were identified. All carcinomas were clustered in the same group by clustering analysis using dysregulated lncRNAs or mRNAs. Four lncRNAs (LINC00959, lnc-FLT3-2:2, lnc-FEZF2-9:2, and lnc-RP11-1035H13.3.1-2:1) identified were significantly dysregulated in further RT-PCR validation. The global "lncRNA score" calculated from the lncRNAs above also differentiated parathyroid carcinomas from adenomas. CONCLUSIONS: LncRNA profiling shows distinct differentially expressed lncRNAs in parathyroid neoplasm. They may play a key role in parathyroid cancer and serve as potential biomarkers to distinguish parathyroid cancers from parathyroid adenomas.

Co-receptor tropism and genetic characteristics of the V3 regions in variants of antiretroviral-naive HIV-1 infected subjects.

Co-receptor tropism has been identified to correlate with HIV-1 transmission and the disease progression in patients. A molecular epidemiology investigation of co-receptor tropism is important for clinical practice and effective control of HIV-1. In this study, we investigated the co-receptor tropism on HIV-1 variants of 85 antiretroviral-naive patients with Geno2pheno algorithm at a false-positive rate of 10%. Our data showed that a majority of the subjects harboured the CCR5-tropic virus (81.2%, 69/85). No significant differences in gender, age, baseline CD4+ T-cell counts and transmission routes were observed between subjects infected with CXCR4-tropic or CCR5-tropic virus. The co-receptor tropism appeared to be associated with the virus genotype; a significantly more CXCR4-use was predicted in CRF01_AE infections whereas all CRF07_BC and CRF08_BC were predicted to use CCR5 co-receptor. Sequences analysis of V3 revealed a higher median net charge in the CXCR4 viruses over CCR5 viruses (4.0 vs. 3.0, P < 0.05). The predicted N-linked glycosylation site between amino acids 6 and 8 in the V3 region was conserved in CCR5 viruses, but not in CXCR4 viruses. Besides, variable crown motifs were observed in both CCR5 and CXCR4 viruses, of which the most prevalent motif GPGQ existed in both viral tropism and almost all genotypes identified in this study except subtype B. These findings may offer important implications for clinical practice and enhance our understanding of HIV-1 biology.

Understanding electro-catalysis by using density functional theory.

The rapid development of catalysts requires a deep understanding of catalytic mechanisms. Since the experimental results have fallen short of the expectation of the optimal catalyst, the density functional theory (DFT) can provide invaluable mechanistic insights and predict promising catalysts. In this perspective, we briefly summarized the advantages of DFT in atomic and electronic structures for understanding electro-catalysis. Some achievements of DFT calculations were reviewed through some examples of the considered catalytic reactions (hydrogen evolution reaction, oxygen reduction reaction, nitrogen reduction reaction, and CO2 reduction reaction). Finally, we highlighted and analyzed the opportunities and challenges in DFT calculations used for electro-catalysis. On the road towards an optimal catalyst, the design of catalysts fused with DFT calculations promises rapid advances in the coming years.

Phase-shifting differential interference contrast microscope with Savart shear prism and rotatable analyser.

A novel differential interference contrast microscope (DICM) is proposed in this research. It is constituted by inserting a Savart shear prism between the objective and sample of a polarising microscope having a rotatable analyser as the phase-shifter, and it is with the ability to enhance image contrast using the principle of shearing interferometry. This letter is to introduce the configuration, interpret the interference patterns and present the experimental setup of the DICM. In addition, this letter is to display the experimental results from the uses of the setup; the results demonstrate the validity and ability of the DICM.

The changing HIV-1 genetic characteristics and transmitted drug resistance among recently infected population in Yunnan, China.

Multiple human immunodeficiency virus (HIV)-1 genotypes in China were first discovered in Yunnan Province before disseminating throughout the country. As the HIV-1 epidemic continues to expand in Yunnan, genetic characteristics and transmitted drug resistance (TDR) should be further investigated among the recently infected population. Among 2828 HIV-positive samples newly reported in the first quarter of 2014, 347 were identified as recent infections with BED-captured enzyme immunoassay (CEIA). Of them, 291 were successfully genotyped and identified as circulating recombinant form (CRF)08_BC (47.4%), unique recombinant forms (URFs) (18.2%), CRF01_AE (15.8%), CRF07_BC (14.4%), subtype C (2.7%), CRF55_01B (0.7%), subtype B (0.3%) and CRF64_BC (0.3%). CRF08_BC and CRF01_AE were the predominant genotypes among heterosexual and homosexual infections, respectively. CRF08_BC, URFs, CRF01_AE and CRF07_BC expanded with higher prevalence in central and eastern Yunnan. The recent common ancestor of CRF01_AE, CRF07_BC and CRF08_BC dated back to 1983.1, 1992.1 and 1989.5, respectively. The effective population sizes (EPS) for CRF01_AE and CRF07_BC increased exponentially during 1991-1999 and 1994-1999, respectively. The EPS for CRF08_BC underwent two exponential growth phases in 1994-1998 and 2001-2002. Lastly, TDR-associated mutations were identified in 1.8% of individuals. These findings not only enhance our understanding of HIV-1 evolution in Yunnan but also have implications for vaccine design and patient management strategies.

[A study of the relations of foramen rotundum structure direction and the approach of percutaneous puncturing of radiofrequency thermocoagulation for treating V2 of primary trigeminal neuralgia].

Objective: To investigate the relations of foramen rotundum structure direction and surrounding structure systematically in order to choose the best approach of percutaneous puncturing of radiofrequency thermocoagulation for treating V2 of primary trigeminal neuralgia. Methods: A total of 122 patients with V2 of primary trigeminal neuralgia for radiofrequency thermocoagulation were enrolled from August 2012 to May 2017 at the First Hospital of Jiaxing. CT scan images were observed retrospectively, to find the inside and outside of the foramen rotundum. The direction of foramen rotundum were recorded and the best approach of puncturing were analyzed. Results: The images were divided into four quadrants with the semi - coronal CT scan plane of the lower margin of the zygomatic arch and the outer edge of foramen rotundum for horizontal axis, and the sagittal plane for the vertical axis. In 122 cases, foramen rotundum direction in outer upper quadrant were 77 cases(63.1%), and in outer under quadrant were 22 cases(18.0%), and in inner upper quadrant were 19 cases(15.6%), and in inner under quadrant were 4 cases(3.3%). Conclusion: The most common foramen rotundum direction is in outer upper quadrant, so the best approach of percutaneous puncturing of radiofrequency thermocoagulation for treating V2 of primary trigeminal neuralgia is the upper side against zygomatic and the inner side against the wall of maxillary sinus.

[Computer Aided Diagnosis system of breast ultrasound based on support vector machine: a clinical analysis].

Objective: To investigate the value of based on support vector machine (SVM) breast ultrasonography technology of Computer-Assisted diagnosis (CAD) for differential diagnosis of benign and malignant breast masses. Methods: Total of 143 patients who had 151 breast masses were collected in Fujian Maternity and Children Health Hospital or The Fist Affiliated Hospital of Fujian Medical University from June 2014 to December 2015. Based on pathological results as the gold standard, the diagnostic efficiency of CAD and ultrasonography were compared. Results: The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of ultrasonography were 80.1%, 71.0%, 76.8%, 80.0% and 72.1%, respectively. And the sensitivity, specificity, accuracy, positive predictive value and negative predictive value of CAD were 96.6%, 90.3%, 94.0%, 93.5% and 94.9%, respectively. The specificity, accuracy, positive predictive value and negative predictive of CAD technology were significantly higher than those of ultrasonography (P<0.05). The area under the receiver operator characteristic curve of ultrasonography and CAD were 0.759 and 0.935 respectively, with significant difference (P<0.05). Conclusions: CAD has a higher specificity and accuracy rate than ultrasonography in the diagnosis of breast masses. It could help to differentiate benign from malignant breast masses.

[Clinical significance of drug resistance-associated mutations in treatment of hepatitis C with direct-acting antiviral agents].

Direct-acting antiviral agents (DAAs) achieve a high sustained virologic response rate in the treatment of chronic hepatitis C virus infection. However, drug resistance-associated mutations play an important role in treatment failure and have attracted more and more attention. This article elaborates on the clinical significance of drug resistance-associated mutations from the aspects of their definition, association with genotype, known drug resistance-associated mutations and their prevalence rates, the impact of drug resistance-associated mutations on treatment naive and treatment-experienced patients, and the role of clinical detection, in order to provide a reference for clinical regimens with DAAs and help to achieve higher sustained virologic response rates.

Inhibition of subcutaneously implanted human pituitary tumor cells in nude mice by LRIG1.

The aim of this study was to explore the inhibition of subcutaneously implanted human pituitary tumor cells in nude mice by LRIG1 and its mechanism. For this study, athymic nude mice were injected with either normal pituitary tumor RC-4B/C cells or LRIG1-transfected RC-4B/C cells. We then calculated the volume inhibition rate of the tumors, as well as the apoptosis index of tumor cells and the expression of Ras, Raf, AKt, and ERK mRNA in tumor cells. Tumor cell morphological and structural changes were also observed under electron microscope. Our data showed that subcutaneous tumor growth was slowed or even halted in LRIG1-transfected tumors. The tumor volumes were significantly different between the two groups of mice (χ2 = 2.14, P < 0.05). The tumor apoptosis index was found to be 8.72% in the control group and 39.7% in LRIG1-transfected mice (χ2 = 7.59, P < 0.05). The levels of Ras, Raf, and AKt mRNA in LRIG1-transfected RC-4B/C cells were significantly reduced after transfection (P < 0.01). Transfected subcutaneous tumor cells appeared to be in early or late apoptosis under an electron microscope, while only a few subcutaneous tumor cells appeared to be undergoing apoptosis in the control group. In conclusion, the LRIG1 gene is able to inhibit proliferation and promote apoptosis in subcutaneously implanted human pituitary tumors in nude mice. The mechanism of LRIG1 may involve the inhibition of the PI3K/ Akt and Ras/Raf/ERK signal transduction pathways.

A diagnostic kit for the enteroviruses Coxsackie A6 and A10.

Recently, there has been an upward trend in the occurrence of hand-foot-mouth disease, which is correlated with Coxsackie A6 and A10 infections. Although two separate diagnostic reagents are available for these two viral strains, the protocol and diagnosis efficiency still need to be improved. More importantly, as co-infection with these viruses is common, the development of a single test kit that can diagnose both viruses would be most beneficial for clinical practice. In our study, specific primers targeting viral nucleic acids were designed and modified. Viral nucleic acids were extracted from fecal or throat swab samples by ultrasonic rupture and silicon membrane purification. The consistency, specificity, and sensitivity of the tests were further optimized by adjusting the polymerase chain reaction (PCR) conditions. The efficiency of viral nucleic acid extraction was significantly enhanced by the ultrasonic rupture and silicon membrane elution approach. Specific amplifications of both viral nucleic acids were achieved using modified primers. The optimal conditions for PCR were also determined (60°C for 30 min and 95°C for 2 min, followed by 40 cycles of denaturation for 30 s at 95°C, annealing for 30 s at 60°C, and elongation for 50 s at 72°C). Amplified products were confirmed as viral specific nucleotides by agarose gel electrophoresis and sequencing. The minimal nucleic acid concentration required for detection was 0.2 ng/L, which was adequate to yield satisfactory specificity and consistency. This novel diagnostic method has many advantages, including rapid protocols and accurate results, and can be promoted for large-scale clinical trials.

Phylogenetic analysis of the Mongolian gerbil (Meriones unguiculatus) from China based on mitochondrial genome.

Meriones unguiculatus (Gerbillinae, Rodentia) is widely used as an animal model of human disease. Here, we provide the first report of the complete mitochondrial genome sequence of M. unguiculatus (GenBank accession Nos. KF425526 and NC_023263). The sequence contained the conserved vertebrate pattern of 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and 1 major noncoding region. We identified one extended termination-associated sequence and one conserved sequence block in the non-coding region. The putative origin of replication for the light strand (OL) was 35 bp long. The OL stem and adjacent sequences were highly conserved, but the loop region differed from those of other rodent species. Base composition and codon usage of the 13 protein-coding genes in M. unguiculatus were compared with those of 23 rodent species with previously sequenced mitochondrial genomes. An A+T content of 63.0% was present in M. unguiculatus; this is similar to the Murinae average (62.4 ± 0.8%) and falls between the average for Mus musculus (63.1 ± 0.1%) and Rattus sp (61.7 ± 0.4%). The AT and GC skew values of M. unguiculatus were 0.035 and -0.28, respectively, similar to those of Cricetinae species (0.057 ± 0.05 and -0.31 ± 0.05). The codon families exhibited similar abundance in all 24 species. Analysis of phylogenetic relationships with 23 other rodent species using neighbor-joining and maximum likelihood protocols and the 12 protein-coding regions on the H strand showed that M. unguiculatus should be classified as genus Meriones, sub-family Gerbillinae, family Muridae.

[Clinical characteristics and prognosis of concurrent positive t(14; 18) and myc gene rearrangement in diffuse large B cell lymphoma].

OBJECTIVE: To study the incidence of positive t(14; 18) and myc gene rearrangement, and the clinical features and prognosis of concurrent positive t(14; 18) and myc gene rearrangement "double-hit lymphoma" (DHL) in diffuse large B cell lymphoma. METHODS: The positive t(14; 18) and myc gene rearrangement in 106 cases of DLBCL were analyzed using interphase fluorescent in situ hybridization (FISH) technique. The expression of myc and bcl-2 proteins was determined by immunohistochemistry. The relationship of positive t(14; 18) and myc gene rearrangement with clinical features, pathogenesis and prognosis for the patients was analyzed. SPSS 16.0 software was used for statistical analysis. RESULTS: Among the 106 cases, there were 27 (25.5%) cases with positive t(14; 18) and 13 (12.3%) cases with myc gene rearrangement, and 7 cases (6.6%) of DLBCL with concurrent t(14; 18)-positive and myc gene rearrangement. A relationship was observed between positive t(14; 18) and myc gene rearrangement (P=0.019). The follow-up data showed that the 7 DHL patients were in age of 52-84 years, the International Prognostic Index (IPI) scores were 3 in two cases, 4 in four cases and 5 in one case, and the ECOG scores were 3 in all the 7 cases. Four patients had bone marrow involvement and were combined with leukemia. The survival time ranged from 0.5 to 6 months, with a median survival of 4 months. The univariate analysis showed that B symptom, Ann Arbor stage, ECOG score, LDH level, IPI score, immunophenotype, bcl-2 protein expression, myc protein expression, and myc gene rearrangement were all associated with poor prognosis (P<0.05 for all). The multivariate analysis using a COX proportional hazard model confirmed that ECOG score, bcl-2 protein expression, myc protein expression, myc gene rearrangement, and immunophenotype were independent prognostic factors affecting survival (P<0.05 for all), among them, the myc gene rearrangement was the strongest prognostic factor (OR=4.337, P<0.001). CONCLUSIONS: "Double-hit" DLBCL is rare and can be mainly identified only by molecular detection. Perhaps positive t(14; 18) and myc gene rearrangement play concurrent role in its "double-hit" pathogenesis. DHL are highly invasive, and most of DHL patients have poor prognosis. Further studies of larger case number are required to determine the pathologic features and the therapeutic strategy of this subgroup.

[Association and interaction of heat shock proteins B1 gene and tumor-suppressor protein p53 gene with chromosome damage levels among coke oven workers].

Objective: To investigate the association and interaction of heat shock proteins B1(HSPB1)gene rs2868371 and tumor-suppressor protein p53(TP53)gene rs1042522 polymorphisms with chromosome damage levels among coke oven workers. Methods: We recruited 1 333 male workers from a state-run coke oven plant in Wuhan in September-October 2010. Among them, 949 who had worked in coke oven workplaces, including auxiliary facilities and bottom, side, and top ovens, were nominated as coke oven workers(i.e., exposed), and 384 administrative or medical staff whose workplaces were offices were used as controls. General characteristics and 5 ml of venous blood were collected from each participant. The plasma concentrations of benzo[a]pyrene-diolepoxide(BPDE)-albumin adducts and the lymphocytic micronucleus(MN)frequencies for each individual were detected by ELISA and cytokinesis-block micronucleus assay, respectively. Gene polymorphisms were genotyped using TaqMan assays via quantitative PCR(ABI Prism 7900HT), and the corresponding frequency ratios(FR)with 95% confidence intervals(CI)were computed for all assays. Results: In the exposed group, the MN frequencies were higher in HSPB1 rs2868371 GC, CC, and GC+ CC genotype carriers((3.88 ± 2.88)‰,(4.00 ± 2.66)‰, and(3.91 ± 2.83)‰, respectively)than in rs2868371 GG genotype carriers((3.52±2.67)‰; FR=1.10, 1.13, and 1.11; 95% CI: 1.02-1.19, 1.02-1.25, and 1.03-1.19, respectively), and the HSPB1 rs2868371C allele was associated with increased MN frequency(Ptrend=0.006). Further, in the exposed group, the MN frequencies were lower in TP53 rs1042522 CG and CG+GG genotype carriers((3.63±2.61)‰ and(3.66±2.61)‰, respectively)than in TP53 rs1042522 CC genotype carriers(3.95±3.06)‰(FR=0.87 and 0.90; 95% CI: 0.83-0.96 and 0.84-0.97, respectively). The effect of gene-gene interaction between HSPB1, rs2868371, and TP53 rs1042522 on MN frequency was significant among coke oven workers(P=0.001). Further stratified analyses showed that the effects of the HSPB1 rs2868371C allele in increasing MN frequencies were robust in subjects aged >40 years(FR=1.07, 95% CI: 1.01-1.12), those working >20 years(FR=1.08, 95% CI: 1.02-1.14), those with BMI ≤24 kg/m2(FR=1.07, 95% CI: 1.01-1.13), drinkers(FR=1.09, 95% CI: 1.02-1.16), and workers with higher BPDE-albumin adduct levels(FR=1.07, 95% CI: 1.01-1.13)(Ptrend=0.023, 0.013, 0.029, and 0.020, respectively). The decreasing effect of the TP53 rs1042522 G allele on MN frequencies was robust in subjects aged >40 years(FR=0.94, 95% CI: 0.89-0.99), those with BMI ≤24 kg/m2(FR=0.94, 95% CI: 0.88-0.99), and drinkers(FR=0.94, 95% CI: 0.88-1.00)(Ptrend=0.031, 0.023, and 0.038, respectively). In addition, there were interactions between HSPB1 rs2868371 and age and between HSPB1 rs2868371 and working years in terms of MN frequency(P=0.030 and 0.013, respectively). Conclusion: In coke oven workers, the HSPB1 rs2868371 C and TP53 rs1042522 G alleles were associated with increased and decreased chromosome damage levels, respectively, and their interaction effect on chromosome damage levels was significant.

[A new challenge in clinical practice: resistance to directly acting antivirals in hepatitis C treatment].

Directly acting antivirals (DAAs) is a major treatment of hepatitis C virus (HCV) overseas. But DAAs resistance is getting more and more clinicians' attention. DAAs have not been approved in China to date, even though some of them are in clinical trials. However, a good knowledge of DAAs resistance is important on optimizing HCV treatment regimens, increasing sustained virological response (SVR) and decreasing treatment failure in clinical. In this review, DAAs resistance mechanism and virologic barrier to resistance, the prevalence of pre-existing DAAs resistance-associated variants (RAVs), the impact of RAVs on treatment outcome, the options of treatment regimens after resistance and drug resistance testing are discussed, hoping to provide some help for DAAs' standardized treatment in China in the future.

[Role of glucose-6-phosphate dehydrogenase in hepatitis B virus replication and its possible mechanism of action].

OBJECTIVE: To investigate the role of glucose-6-phosphate dehydrogenase (G6PD) in hepatitis B virus (HBV) replication and its possible mechanism of action. METHODS: Tissue microarray, quantitative real-time PCR, and Western blot were performed to analyze the differences in G6PD expression levels in the HBV-positive and HBV-negative liver tissues, HepG2.2.15 cells, and HepG2 cells. The siRNA transfection technique was used to knock down G6PD gene in HepG2.2.15 cells for 48 hours. Chemiluminescence was used for HBsAg and HBeAg quantification in supernatant, and quantitative real-time PCR was used to measure HBV DNA, type I interferon (IFN), and downstream IFN-stimulated genes. The t-test was used for comparison between groups. RESULTS: G6PD expression was significantly upregulated in the HBV-positive liver tissues and cells compared with HBV-negative liver tissues and cells, and the stain intensity and immunohistochemical scores were 89.69±54.92 and 31.90±18.62, respectively (P < 0.05). After G6PD expression in HepG2.2.15 cells was interfered by siRNA, the quantitative levels of HBV DNA, HBsAg, and HBeAg in supernatant were reduced significantly, and the mRNA expression levels of IFNα1, IFNß1, and five downstream IFN-stimulated genes (OAS1, ISG15, OAS3, EIF2α, and PKR) increased significantly (all P < 0.05). CONCLUSION: G6PD plays a vital role in HBV replication, and its mechanism of action in regulating HBV replication may be related to type I IFN signaling pathway.