[Applications and challenges of pathomics technique in the management of hepatocellular carcinoma].

The incidence and mortality rate of hepatocellular carcinoma rank among the top of all cancer types,seriously threatening the life and health of human beings. In recent years,the rapid development of artificial intelligence and the deepening of the concept of precision medicine have led to a boom in interdisciplinary research. Pathomics,as an emerging omics technology driven by artificial intelligence,can mine massive information from high-resolution whole slide images,and shows broad application prospects in the diagnosis,treatment and prognosis assessment of hepatocellular carcinoma. However, pathomics research in hepatocellular carcinoma is still in its infancy, and its research patterns and clinical applications still face several controversies and challenges, including data security, ethics, and "black box" issues. Future research should focus on conducting prospective studies, integrating multimodal data, improving computational technologies, and establishing professional standards to promote the high-quality development of pathomics technology in both clinical and basic research of hepatocellular carcinoma.

[Genotype and genetic characteristics of human respiratory syncytial virus circulating in Quanzhou, 2018-2019].

Objective: To study the prevalence and genetic characteristics of human respiratory syncytial virus (HRSV) in Quanzhou city, from 2018 to 2019. Methods: A total of 141 throat swabs were collected from children patients of lower respiratory tract infection in Quanzhou children Hospital, Fujian Province from November 2018 to May 2019. RT-PCR was used to amplify the 3 'end of G gene HRSV. Sequencer 5.0 and MEGA5.05 softwares were used for sequence editing, phylogenetic tree construction and genotyping analysis. Results: Twenty-five samples were positive for HRSV. Seventeen samples succeeded to obtain the target gene, including 13 of HRSVA and 4 of HRSVB. Two genotypes were identified: ON1 genotype (13 samples, HRSVA) and BA9 genotype (4 samples, HRSVB). Five strains of ON1 genotype sequences were clustered with the ON1 sequences prevalent in Beijing, Changchun and Zhejiang from 2012 to 2015 (cluster1); one strain (FJ19-02) was clustered with the sequences of ON1 genotype circulating in many regions of China from 2012 to 2015 (cluster2); Seven strains were clustered independently (cluster FJ). FJ18-02, FJ19-14 and FJ19-15 of HRSVB were clustered with the BA9 genotype sequences prevalent in Changchun, Jilin Province in 2015, while FJ19-13 was closely related to the BA9 genotype sequences prevalent in Guangzhou and Zhejiang Province in 2013. Both the ON1 and BA9 genotypes showed variations of nucleotide and amino acid in 72 and 60 insertion segments. Amino acid mutation (H266L) only occurred among the sequence of cluster-FJ, and the mutations of H261Q and Q265L only appeared in strain FJ19-13. Conclusion: BA9 and ON1 genotypes were prevalent in Quanzhou city, from 2018 to 2019. Cluster-FJ was a newly discovered independent transmission chain, which may continue to circulate in local Quanzhou area.

[Application of cell transfer technique in the diagnosis of fine needle aspiration cytology].

Objective: To study the application of cell transfer technology to solve the problem of the limited number of fine needle aspiration cytology (FNAC) smears for various immunocytochemistry (ICC) staining and other auxiliary tests, and to enhance accurate cytological diagnosis. Methods: Thirty-four cases of FNAC smears from January 2020 to April 2020 in the Department of Pathology of Beijing Hospital were collected for investigation of the cell transfer technique. The materials in the most cell smear were divided and transferred to several glass slides. After de-staining, the recipient slides were stained with EnVision ICC. The technique was validated by comparing the consistency of the ICC of transferred cell smears and the corresponding immunohistochemical (IHC) staining on biopsies. Results: There were a total of 180 cell transfer slides from 34 cases, of which 174 had the same cell morphology, size and structure as the original smears, with the success rate of cell transfer of 96.7% (174/180). Totally 174 ICC stains were performed on the successfully transferred cell smears, of which 153 smears had available corresponding IHC staining of histologic specimens. Of these, 148 showed concordance between ICC staining and the IHC staining. Cells were successfully transferred in 96.7 % (148/153) of the cell sheets, keeping the same morphology and structure as compared to their original smears. The diagnosis of all 34 FNAC cases was the same to that of their corresponding pathology on biopsies with 100 % concordance. Conclusions: The cell transfer technique is a simple and effective way to make full use of diagnostic cells on a cell smear, and is valuable for accurate cytological diagnosis.

[Occurrence and related factors of preterm birth in Guangxi Zhuang Autonomous Region during 2017-2019].

Objective: To investigate the incidence of preterm birth in Guangxi Zhuang Autonomous Region and explore the related factors and their combined effects. Methods: The study subjects were women giving birth to live babies at the monitoring points of critical maternal hospital monitoring system in Guangxi Zhuang Autonomous Region from January 1, 2017 to December 31, 2019. The data of general characteristics (age and marital status), pregnancies (parity, number of previous cesarean delivery, the number of prenatal check and number of fetuses in this pregnancy) and disease conditions (placenta previa, placental abruption, hypertension, diabetes, anemia, and heart disease) were collected, and the incidence of preterm birth were calculated according to the definition of preterm birth set by WHO and China, respectively. Logistic regression model was used to explore the factors associated with premature birth and their combined effects. Results: According to definitions of WHO and China, the cumulative incidence of preterm birth in Guangxi from 2017 to 2019 was 7.45% (16 819/225 727) and 7.34% (16 559/225 727), respectively. Advanced age [≤34 years old as reference, OR (95%CI) of 35-39 and ≥40 years old were 1.36 (1.30-1.42) and 1.61 (1.50-1.74), respectively], unmarried (including divorced or widowed) [OR (95%CI): 1.28 (1.17-1.40)], primiparae [OR (95%CI): 1.34 (1.29-1.40)], previous cesarean section [no previous cesarean section as reference, OR (95%CI) of 1 and ≥2 times of previous cesarean section were 1.30 (1.24-1.36) and 1.85 (1.65-2.08), respectively], antenatal examination<8 [OR (95%CI): 2.72 (2.62-2.81)], multiple pregnancies [OR (95%CI): 15.00 (14.01-16.06)], placenta previa [OR (95%CI): 6.90 (6.35-7.50)], placental abruption [OR (95%CI): 8.18 (7.36-9.10)], gestational hypertension [OR (95%CI): 2.29 (2.17-2.42)], gestational diabetes mellitus [OR (95%CI): 1.43 (1.37-1.49)], anemia [OR (95%CI): 1.10 (1.07-1.14)], and heart diseases [OR (95%CI): 2.98(2.43-3.65)] were all positively correlated with preterm birth. The risk of preterm birth in pregnant women exposed to 1, 2, 3, 4, 5, 6 and ≥7 preterm birth related factors was 1.51, 2.29, 4.49, 9.69, 20.87, 46.88 and 192.11 times that of non-exposed women, respectively (all P values<0.001). Conclusion: Preterm birth is associated with maternal general characteristics, pregnancy and disease status, and the combined effect of preterm birth related factors significantly increases the risk of preterm birth.

[Investigation on the awareness of snoring hazard and prevalence of obstructive sleep apnea among civil servants].

Objective: To investigate the awareness of snoring hazard and prevalence of obstructive sleep apnea (OSA) among civil servants. Methods: A cross-sectional survey was conducted to investigate the awareness of snoring hazards among in-service civil servants who had annual medical examination in a Guangdong provincial institution from September to November 2017. The high-risk group for OSA was screened and diagnosed by sleep monitoring. Results: 1 036 of 1 241 civil servants were enrolled in the study for integral data. 60.1% (623/1 036) of the subjects realized that snoring was harmful to health. The most common source to develop OSA awareness was network (59.6%, 371/623), followed by television (48.0%), relatives and friends (46.6%), newspaper (44.5%) and radio (18.9%). The awareness rate of snoring consequences was as follows: decreased sleep quality (71.9%, 448/623), sudden death (52.2%), daytime sleepiness (44.3%), cardiovascular and cerebrovascular diseases (42.9%), hypertension (24.4%) and sexual dysfunction (16.7%). 22.0% (228 / 1 036) of the cases were classified into high-risk OSA. The prevalence of OSA among high-risk group was 46.05%(105/228)and only 0.9% (2/228) of them had been diagnosed with OSA. Conclusion: Civil servants had awareness of snoring hazard to a certain extent. Among civil servants classified into high-risk OSA, the OSA perveance was high but the rate of diagnosis and treatment was very low.

[Long non-coding RNA RAB11B-AS1 prevents osteosarcoma proliferation via its sense gene RAB11B].

Objective: To detect the expression of lncRNA RAB11B-AS1 in osteosarcoma and investigate its role in osteosarcoma cells proliferation and the responsible mechanisms. Methods: Osteosarcoma and corresponding adjacent normal tissues were collected from 24 patients subjected to operations from October 2015 to October 2017 in the Third Affiliated Hospital of Southern Medical University.RAB11B-AS1 expression was detected in osteosarcoma specimens by quantitative real-time polymerase chain reaction (qRT-PCR). Lentiviral vectors that stably over-expressing RAB11B-AS1 were constructed and transfected into U2OS osteosarcoma cell line.The effect of RAB11B-AS1 on osteosarcoma cell proliferation and apoptosis was investigated by cell counting kit (CCK-8) assay and flow cytometry.U2OS osteosarcoma xenograft model of nude mice was established to observe the effect of RAB11B-AS1 on xenograft growth in mice, and the role of RAB11B-AS1 in proliferation and apoptosis of osteosarcoma cells was investigated by immunohistochemistry and TUNEL staining of osteosarcoma slices.The relationship between RAB11B-AS1 and RAB11B was explored using luciferase reporter assay.The data were compared with t test between the two groups. Results: Expression of RAB11B-AS1 was significantly down-regulated in osteosarcoma (0.010±0.015) versus their paired non-neoplastic tissues (0.022±0.030) (t=2.117, P=0.045). Up-regulation of RAB11B-AS1 resulted in decreased proliferative rate of U2OS cells (F=15.659, P<0.001). The ratios of cells in G0-G1 phase, S phase, G2-M phase were 62.6%±6.3%, 21.4%±2.2%, 16.3%±1.6% respectively in RAB11B-AS1 up-regulated group versus 59.4%±5.9%, 25.9%±2.6%, 15.5%±1.1% respectively in control group, and cell ratio in G0-G1 and S phase were increased significantly by RAB11B-AS1 up-regulation (t=17.124, 17.321, both P<0.05). Apoptosis rate was significantly elevated in RAB11B-AS1 over-expressed cells (12.7%±1.3%) when compared with that in control (10.3%±1.0%)(t=17.321, P=0.003). Mice transplanted with osteosarcoma cells that overexpressed RAB11B-AS1 exhibited lower growth rate of tumor (F=8.798, P=0.009). Mechanistically, RAB11B-AS1 expression correlated negatively with RAB11B expression (r=-0.356, P=0.044). Conclusions: lncRNA RAB11B-AS1 expression is down-regulated significantly in osteosarcoma tissues.RAB11B-AS1 may suppress the progression of osteosarcoma via down-regulating RAB11B.

[Determination of iodine content in the left ventricular myocardium of healthy subjects using dual-source dual-energy CT myocardial first perfusion imaging: a preliminary study].

Objective: To analyze the normal value of the iodine content in the left ventricular myocardium of healthy subjects and to observe if there is a segmental differences on iodine distribution by using the second generation dual-source dual-energy computed tomography myocardial first perfusion imaging. Methods: In this retrospective study, 42 healthy subjects, who admitted to our department between January to June 2016, with normal second generation dual-source dual-energy computed tomography and coronary CT angioghphy (CTA), electrocardiogram (ECG) results, normal cardiac, hepatic, renal function, normal myocardial enzymes results were enrolled, data from 38 out of 42 subjects with satisfactory image quality were analyzed using Siemens Dual Energy-Heart PBV image processing software.In accordance with the standards of the American Heart Association myocardial 17 fractionation method, content of iodine was measured at different segmental left ventricular myocardium and aorta (left coronary artery from the opening level). The standardized containing iodine value (nIC) was calculated. Results: The iodine content of left ventricular myocardium in normal subjects was 3.1-7.8 mg/ml.The nIC of myocardium from 1st to 17th segments was 0.28±0.06, 0.31±0.07, 0.30±0.07, 0.30±0.04, 0.28±0.04, 0.29±0.05, 0.29±0.01, 0.30±0.07, 0.31±0.07, 0.27±0.06, 0.28±0.08, 0.28±0.07, 0.29±0.08, 0.31±0.07, 0.27±0.06, 0.29±0.06 and 0.21±0.07, respectively.The nIC of the 17th segment was the lowest and was significantly lower than in other segments (all P<0.05), the nIC was similar among the rest 16 segments (all P>0.05). Conclusion: The normal iodine content range in left ventricle myocardium is 3.1-7.8 mg/ml, and the lowest iodine content is detected in the apex and which is significantly lower than the other left ventricular segments.

[Intervention of edaravone against renal injury induced by acute paraquat poisoning in rats].

Objective: To investigate the dynamic change of paraquant-induced kidney injury in rats and the protective effect of edaravone. Methods: Eighty SD rats were randomly divided into 4 groups: the normal control group, paraquat poisoning group, edaravone treatment group and edaravone control group. The normal control group of 8 rats were given 1 ml of 0.9% sodium chloride through the abdominal cavity, and the same amount of fluid into the abdominal cavity after 30 minutes. The paraquat poisoning group of 24 rats were given 1 ml of paraquat solution (20 mg/kg) through the abdominal cavity to build poisoning models, and the same amount of 0.9% sodium chloride was injected into the abdominal cavity after 30 minutes. The edaravone treatment group of 24 rats were given edaravone (5 mg/kg) through the abdominal cavity after 30 minutes when the poisoning models were set up. The edaravone control group of 24 rats were given 1 ml of 0.9% sodium chloride through the abdominal cavity, and edaravone (5 mg/kg) was injected into the abdominal cavity after 30 minutes. In addition to the normal control group, the other groups processed 1 times a day to mantain 7 d. On 1, 3, 7, 21 d several rats in each group were excuted and the kidney tissue and serum samples were collected, then each pathological changes of the kidney were observed with light microscopy. Serum creatinine, KIM-1, NGAL were measured by ELISA, the expression of HSP70 protein in kidney were observed with immunohistochemical staining. Results: The pathological examination reveald that the damage of kidney tissue in the paraquat group was the most serious on 3 d, and the damage was consistently alleviated in edaravone treatment group at the same time, renal fibrosisn was unseen in each group until 21 d. Compared with normal control group, there was no statistically significant in edaravone control group (P>0.05) . The KIM-1 in blood and kidney in paraquat poisoning group were markedly increased in 1 d (P<0.05) . The NGAL in blood and creatinine were markedly increased in d7 (P<0.05) . The NGAL in kidney increased over time, but had no statistically difference with the control group (P>0.05) .Compared with paraquat poisoning group, the serum creatinine, KIM-1 in blood and kidney, the KIM-1 in kidney had decreased significantly in edaravone treatment group (P<0.05) . The NGAL in kidney has no statistically significant compared with the poisoning group (P>0.05) . HSP70 expression of kidney tissue in edaravone treatment group had significantly increased in d3 compared with the paraquat poisoning group (P<0.05) . Conclusion: Edaravone can prompt a significant rise of HSP70 in kidney tissue, reduce KIM-1 and NGAL levels, and play a protective role in kidney injury of acute paraquat poisoning.

Development of expressed sequence tag-simple sequence repeat markers for genetic characterization and population structure analysis of Praxelis clematidea (Asteraceae).

Invasive plants tend to spread aggressively in new habitats and an understanding of their genetic diversity and population structure is useful for their management. In this study, expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed for the invasive plant species Praxelis clematidea (Asteraceae) from 5548 Stevia rebaudiana (Asteraceae) expressed sequence tags (ESTs). A total of 133 microsatellite-containing ESTs (2.4%) were identified, of which 56 (42.1%) were hexanucleotide repeat motifs and 50 (37.6%) were trinucleotide repeat motifs. Of the 24 primer pairs designed from these 133 ESTs, 7 (29.2%) resulted in significant polymorphisms. The number of alleles per locus ranged from 5 to 9. The relatively high genetic diversity (H = 0.2667, I = 0.4212, and P = 100%) of P. clematidea was related to high gene flow (Nm = 1.4996) among populations. The coefficient of population differentiation (GST = 0.2500) indicated that most genetic variation occurred within populations. A Mantel test suggested that there was significant correlation between genetic distance and geographical distribution (r = 0.3192, P = 0.012). These results further support the transferability of EST-SSR markers between closely related genera of the same family.

Overexpression of OsOSM1 Enhances Resistance to Rice Sheath Blight.

Sheath blight (SB), caused by Rhizoctonia solani, is one of the most destructive rice diseases worldwide. It has been difficult to generate SB-resistant varieties through conventional breeding because of the quantitative nature of rice resistance to SB. In this study, we found that overexpression of the OsOSM1 gene, encoding an osmotin protein belonging to the pathogenesis-related protein 5 family, is able to improve rice resistance to SB in field tests. Although there are two osmotin genes in rice, OsOSM1 is the one mainly expressed in leaf sheath at the booting stage, coinciding with the critical stage of SB development in the field. In addition, OsOSM1 expression is strongly induced by R. solani in SB-resistant rice variety YSBR1 but not in susceptible varieties, suggesting its involvement in SB resistance. Overexpression of OsOSM1 (OsOSM1ox) in susceptible variety Xudao 3 significantly increases resistance to SB in transgenic rice. The OsOSM1 mRNA levels in different transgenic lines are found to be positively correlated with their SB resistance levels. Intriguingly, although extremely high levels of OsOSM1 were detrimental to rice development, appropriately elevated levels of OsSOM1 were obtained that enhanced rice SB resistance without affecting rice development or grain yield. The OsSOM1 protein is localized on plasma membrane. OsOSM1 is upregulated by jasmonic acid (JA); furthermore, JA-responsive marker genes are induced in OsOSM1ox lines. These results suggest that the activation of JA signaling pathway may account for the increased resistance in transgenic OsOSM1ox lines. Taken together, our results demonstrate that OsOSM1 plays an important role in defense against rice SB disease and provides a new target for engineering resistance to SB.

Overexpression of OsPGIP1 Enhances Rice Resistance to Sheath Blight.

Rice sheath blight (SB), caused by necrotrophic pathogen Rhizoctonia solani, is one of the most destructive rice diseases, and no major resistance genes are available. Polygalacturonase-inhibiting proteins (PGIP) are extracellular leucine-rich repeat proteins and play important roles in plant defense against different pathogenic fungi by counteracting secreted fungal polygalacturonases (PG). However, the role of PGIP in conferring resistance to rice SB remains to be thoroughly investigated. Here, we showed that OsPGIP1 is capable of inhibiting PG derived from R. solani. Our real-time reverse-transcription polymerase chain reaction results indicated that resistant rice 'YSBR1' and 'Jasmine 85' express significantly higher levels of OsPGIP1 than susceptible 'Lemont'. Our results also show that OsPGIP1 is most highly expressed at the late tillering stage in the sheath of YSBR1, coinciding with the critical stage of SB development in field. More importantly, the OsPGIP1 level is highly elevated by inoculation with R. solani in resistant cultivars but not in susceptible Lemont. Overexpression of OsPGIP1 significantly increased rice resistance to SB and inhibited tissue degradation caused by R. solani-secreted PG. Furthermore, OsPGIP1 overexpression did not affect rice agronomic traits or yield components. Together, our results not only demonstrate the important role of OsPGIP1 in combatting the rice SB disease but also provide a new avenue to the improvement of rice SB resistance by manipulating an endogenous gene.

[Interferon α sensitizes human osteosarcoma cells to doxorubicin-induced apoptosis through p53-dependent pathway].

Objective: To investigate the effect of IFNα on doxorubicin-induced apoptosis in human osteosarcoma cells with its molecular mechanisms to provide evidences for improving the treatment of osteosarcoma. Methods: Osteosacoma U2OS and MG63 cells were treated with IFNα and Doxorubicin, alone or in combination, for 72 h . Cytotoxicity was determined with MTT. Apoptosis was evaluated through fluorescence-activated cell sorting, Hoechst33258 staining and DNA ladder assay. The expression of p53, Bax, Bcl-2, Mdm2, p21, caspase-3 and PARP was determined with Western blot. siRNA interference was used to silence p53. Results: IFNα treatment for 72 h did not induce cytotoxicity but greatly enhanced doxorubicin-induced cytotoxicity and apoptosis in p53-wild U2OS cells but not p53-mutant MG63 cells. Compared with other groups, the combination of IFNα and doxorubicin induced more obvious apoptotic morphological changes and DNA ladder. IFNα did not alter the expression of the indicate genes. The expression of p53, Bax, Mdm2 and p21 was up-regulated by doxorubicin and further increased in response to combination. The expression of Bcl-2 was down-regulated by doxorubicin and further decreased in response to combination.There were no differences among groups in MG63 cells. The expression of p53 was effectively blocked by p53-siRNA in U2OS cells. The p53 silencing greatly reduced the cytotoxicity mediated by combination for 72 h, compared with non- and control-siRNA groups. The activation of caspase-3 and PARP mediated by combination was largely suppressed by p53 silence. Conclusion: IFNα sensitizes human osteosarcoma cells to doxorubicin-induced apoptosis through p53-dependent pathway. The combination of IFNα and traditional chemotherapy can be used in osteosarcoma treatment.

A novel surface modification scheme for ITO nanocrystals by acetylene: a combined experimental and DFT study.

Many applications of Sn-doped indium oxide (ITO) films in organic electronics require appropriate surface modifications of ITO nanocrystals with small organic molecules, such as silanes, phosophonic acids and carboxylic acids, to improve interfacial contacts and charge transfer. Here, we propose a new surface modification strategy via adsorption of acetylene molecules on an oxygen-terminated ITO(100) surface using a slab crystalline model to represent the nanocrystal surface. The adsorption was first studied using density functional theory. It was found that the chemisorption of C2H2 on two types of surface oxygen dimers is highly exothermic with the calculated adsorption energies of 3.80 eV and 5.19 eV, respectively. Electron population analysis reveals the origin of the strong interaction between the adsorbate and the ITO(100) surface. Experimental studies on the synthesized ITO nanocrystals using X-ray photoelectron spectroscopy and diffuse reflectance infrared Fourier transform spectroscopy confirm the predicted strong adsorption of C2H2 on ITO surfaces.

Drought-tolerant rice germplasm developed from an Oryza officinalis transformation-competent artificial chromosome clone.

Oryza officinalis has proven to be a natural gene reservoir for the improvement of domesticated rice as it carries many desirable traits; however, the transfer of elite genes to cultivated rice by conventional hybridization has been a challenge for rice breeders. In this study, the conserved sequence of plant stress-related NAC transcription factors was selected as a probe to screen the O. officinalis genomic transformation-competent artificial chromosome library by Southern blot; 11 positive transformation-competent artificial chromosome clones were subsequently detected. By Agrobacterium-mediated transformation, an indica rice variety, Huajingxian 74 (HJX74), was transformed with a TAC clone harboring a NAC gene-positive genomic fragment from O. officinalis. Molecular analysis revealed that the O. officinalis genomic fragment was integrated into the genome of HJX74. The transgenic lines exhibited high tolerance to drought stress. Our results demonstrate that the introduction of stress-related transformation-competent artificial chromosome clones, coupled with a transgenic validation approach, is an effective method of transferring agronomically important genes from O. officinalis to cultivated rice.

Molecular identification of hookworms in stray and shelter dogs from Guangzhou city, China using ITS sequences.

Canine hookworm infections are endemic worldwide, with zoonotic transmission representing a potentially significant public health concern. This study aimed to investigate hookworm infection and identify the prevalent species from stray and shelter dogs in Guangzhou city, southern China by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based on internal transcribed spacer (ITS) sequences. From March 2011 to July 2012, fresh faecal samples from a total of 254 dogs were obtained from five locations, namely Conghua, Baiyun, Liwan, Haizhu and Panyu, in Guangzhou. These samples were screened for the presence of hookworm eggs using light microscopy, with an overall prevalence of 29.53% being recorded. The highest prevalence of 45.28% was found in suburban dogs from Conghua compared with lower values recorded in urban dogs in Haizhu (21.43%), Baiyun (18.97%), Panyu (18.18%) and Liwan (15%). The prevalence in stray dogs was significantly higher than that in shelter dogs. PCR-RFLP analysis showed that 57.33% were detected as single hookworm infections with Ancyclostoma caninum, and 22.67% as A. ceylanicum, while 20% were mixed infections. This suggests that high prevalences of both hookworm species in stray and shelter dogs in China pose a potential risk of transmission from pet dogs to humans.

Molecular characteristics of vancomycin-resistant Enterococcus faecium from a tertiary care hospital in Chengdu, China: molecular characteristics of VRE in China.

The aim of this study was to characterize vancomycin-resistant Enterococcus faecium (VREfm) isolates phenotypically and molecularly, and investigate associations between the virulence factors enterococcal surface protein (esp), hyaluronidase (hyl), and collagen adhesin (acm) and colonization/infection. A total of 126 E. faecium [66 VREfm and 60 vancomycin-susceptible (VSEfm)] were collected in West China Hospital. Nine E. faecium isolates (7 VREfm and 2 VSEfm) were selected at random for comparative study in a large region from China. Minimum inhibitory concentrations (MICs) were measured by Etest and agar dilution, vancomycin resistance genes (vanA, vanB, and vanC) and virulence genes (esp, acm, and hyl) were detected by polymerase chain reaction (PCR). Thirty-four VREfm underwent repetitive sequence-based PCR (rep-PCR) and multi-locus sequence typing (MLST). One linezolid-resistant isolate (MIC = 8 µg/ml) was found; none were tigecycline resistant. All 73 VREfm (28 infective strains and 45 intestinal colonizers) had the vanA gene and VanA phenotype. Positivity for esp, hyl, and acm in VREfm was 79.5, 46.6, and 86.3%, respectively, which was higher than in VSEfm (54.8, 27.4, and 56.5%, respectively). Among VSEfm, positivity for acm in isolates from pleural or cerebrospinal fluid (84.6%) was higher than that from blood (32.4%). There were 11 rep-PCR types (similarity >95%) and MLST revealed nine sequence types (STs) among the selected isolates. Most VREfm and all VSEfm belonged to clonal complex 17. A new ST was found, with allele sequence (15, 1, 38, 1, 1, 1, 1). In China, most VREfm seem to belong to the classical nosocomial CC17 clone, and many of them have acquired virulence genes, further strengthening a hospital-adapted type.

Hepatitis B virus X protein activates human hepatic stellate cells through upregulating TGFß1.

We investigated the effects of the hepatitis B virus X gene (HBV X) on the activation of human hepatic stellate cells (HSCs) and the possible mechanisms underlying the pathway. Recombinant plasmid pHBV-X-IRES2-EGFP was constructed and transfected into HL-7702 cells using a lipid-mediated method. Transfected cells were screened by G418, which detected stable expression of the X gene by reverse transcription (RT)-PCR and Western blot analysis, and named L02/x. Cells not subjected to G418-selection were analyzed to confirm the transient expression of the X gene and named L02/48x. Subsequently, L02/x and L02/48x, together with non-HBx-expressing cells, were co-cultured with HSCs in a non-contact transwell system. After 36 h of co-culture, the proliferation and migration of HSCs was detected using different cell counting methods. Finally, the mRNA and protein levels of α-SMA, Col I, and TGFß1 in HSCs were detected by real-time PCR and western blot analysis. RT-PCR and Western blot analysis showed that L02/x and L02/48x cells can express HBV X gene mRNA and protein. Additionally, HSCs co-cultured with L02/x or L02/48x cells showed significantly higher proliferation and migration levels than control groups. Real-time PCR and Western blot analysis showed that the mRNA and protein expressions of α-SMA, Col I, and TGFß1 in HSCs co-cultured with HBx-expressing liver cells were higher than those in control groups. HBx protein activated HSCs in vitro, leading to increased proliferation and migration of HSCs and upregulation of α-SMA and Col I. The TGFß1 gene may be involved in this pathway.

Cloning and sequence analysis of the Blumea balsamifera DC farnesyl diphosphate synthase gene.

Blumea balsamifera DC is a member of the Compositae family and is frequently used as traditional Chinese medicine. Blumea balsamifera is rich in monoterpenes, which possess a variety of pharmacological activities, such as antioxidant, anti-bacteria, and anti-viral activities. Farnesyl diphosphate synthase (FPS) is a key enzyme in the biosynthetic pathway of terpenes, playing an important regulatory role in plant growth, such as resistance and secondary metabolism. Based on the conserved oligo amino acid residues of published FPS genes from other higher plant species, a cDNA sequence, designated BbFPS, was isolated from B. balsamifera DC using polymerase chain reaction. The clones were an average of 1.6 kb and contained an open reading frame that predicted a polypeptide of 342 amino acids with 89.07% identity to FPS from other plants. The deduced amino acid sequence was dominated by hydrophobic regions and contained 2 highly conserved DDxxD motifs that are essential for proper functioning of FPS. Phylogenetic analysis indicated that FPS grouped with other composite families. Prediction of secondary structure and subcellular localization suggested that alpha helices made up 70% of the amino acids of the sequence.

Comparison and Confirmation of Quantitative Trait Loci Conferring Partial Resistance to Rice Sheath Blight on Chromosome 9.

Sheath blight (SB), caused by Rhizoctonia solani, is one of the worst rice (Orzya sativa) diseases worldwide. Resistance to the SB disease in rice is a complex trait controlled by quantitative trait loci (QTLs). Through map integration, we found several previously identified SB resistance (SBR) QTLs reported in inconsistent regions on the long arm of chromosome 9. Five of them were detected on 'Jasmine 85' (J85), 'Minghui 63' (MH63), and 'Lemont' (LMNT) rice and were designated qSB-9J85-1, qSB-9J85-2, qSB-9MH63-1, qSB-9MH63-2, and qSB-9LMNT, respectively, in the present study. To further verify and physically map the five potential SBR QTLs, we introduced these SBR QTLs into a common susceptible variety (LMNT) and developed a few chromosomal segment substitution lines through marker-assisted selection. After artificial inoculation with the SB fungus, we were able to validate qSB-9J85-2 but not the other four SBR QTLs; whereas, on MH63, an SBR QTL designated qSB-9MH63-3 was confirmed in the region defined by markers Y83 and Y91.8 that included qSB-9J85-2, covering approximately 1,235 kb. Both qSB-9J85-2 and qSB-9MH63-3 appeared to be dominant resistance genes and contributed to similar levels to SB resistance, reducing SB disease severity by approximately 1.0 on a 0-to-9 SB disease rating system. After comparing with another confirmed SBR QTL (qSB-9TQ) from 'Teqing' rice (TQ), we conclude that qSB-9J85-2, qSB-9MH63-3, and qSB-9TQ are probably controlled by the same allelic resistance genes. These results will accelerate the utilization of this major SBR QTL and its map-based cloning.