NS2 and NS3 are two post-transcriptional gene silencing suppressors that are encoded by Rice stripe virus. Gene silencing suppressors are always related to the pathogenicity of viruses. In this study, the cDNA of NS2 and NS3 were recombined by overlapping PCR assays, ligated to the RNAi vector, and inserted into the PXQ expression vector using Pst I; the expressed vector was transferred into calluses induced from seeds of the japonica rice cultivar, 'Nipponbare', using an Agrobacterium-mediated method. Thirty-one T0 transgenic plants were selected by G418 screening. PCR and southern blot analyses confirmed that the target gene was transformed into transgenic rice successfully, and different transgenic plants contained various copies of the gene. The disease resistance assay revealed that T0 transgenic rice had a delayed onset of RSV for approximately 10-20 d, and the accumulation of virus in the transgenic plants was reduced by 30%-50%. This was related to the delayed onset of disease.
Disease Resistance ; Oryza ; genetics ; immunology ; virology ; Plant Diseases ; genetics ; immunology ; virology ; Plants, Genetically Modified ; genetics ; immunology ; virology ; RNA Interference ; Tenuivirus ; genetics ; immunology ; Viral Nonstructural Proteins ; genetics ; immunology

Adolescent ; Adult ; Aged ; Female ; Humans ; Joint Dislocations ; physiopathology ; therapy ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Shoulder Joint ; injuries ; physiopathology ; Treatment Outcome ; Young Adult

In vitro gene delivery, polyethyleneimine (PEI) has been described as one of the most efficient nonviral vector. Herein the formation mechanism of PEI/DNA complexes is elucidated. The transition phase of "bead-on-string" structure in the formation of complexes was supposed to exist through spectroscopy, electrophoresis and transmission electron microscopy (TEM) technology. The construction of PEI/DNA complexes is related closely to the characteristics of PEI and DNA plasmid. As well as the dominant electrostatic effects, the nonelectrostatic interactions were thought to be partially responsible for the presence of PEI/DNA complexes even in the high ionic strength. The surface charge of complexes particles increased with the N/P ratio, but the absolute value of zeta potential was lower at the N/P ratio of 8 and 12, perhaps attributed to the use of larger DNA plasmid. As a result, the repulsion between particles was decreased and prone to aggregate to the structure like a clustered grape-string in the solution. Interestingly, contrast to the formation behavior of complexes, the PEI/DNA complexes aggregated primarily due to hydrophobic interactions while electrostatic attractions play a little role in the complexes particles aggregation in different concentrations of salt solutions. Comparable transfection efficiency in HepG2 cells was observed for the Lipofectamine 2000 and PEI/DNA complexes at the N/P ratio of 12, and showed that larger or aggregable complexes could transfect the cells in some different mechanisms.
DNA ; genetics ; Drug Carriers ; Gene Transfer Techniques ; Genetic Therapy ; Polyethyleneimine ; chemistry ; Transfection

The aim of this study was to investigate the mechanism to reverse the drug resistance of leukemia cells in tetrandrine (Tet) alone or in combination with droloxifen (Drol) by using protein chips and to lay the theoretical basis for the clinical applications. Three monoclonal antibodies against P-glycoprotein (P-gp), the multidrug resistance-associated protein (MRP1) and the breast cancer resistance protein (BCRP) were immobilized onto the agarose gel film-coated glass slides. Protein chips were prepared respectively from K562/A02 cells cultured for 12, 24 and 48 hours with Tet alone or in combination with Drol. The results showed that Tet alone or in combination with Drol could decrease only the expression of P-gp in a time-dependent manner, the effect for 48 hours as follows: Tet + Drol 82.620 +/- 3.227; Tet alone 86.440 +/- 2.906; Drol alone 87.230 +/- 2.049; control 93.670 +/- 2.748 (P < 0.05). However, down-regulation of P-gp by K562/A02 cells cultured with Tet alone or in combination with Drol began at 24 hours (Tet + Drol 85.270 +/- 3.095; control 93.670 +/- 2.748, P < 0.05). The results were coincident with that of FCM. It is concluded that Tet and Drol can downregulate the expression of P-gp in the time-dependent way. There is a significant difference between Tet alone and Tet combined with Drol at 24 hours (P < 0.05). The expression of MRP1 and BCRP are not closely correlated with the reversal mechanism of Tet and Drol, and which may be involved in the mechanism of this combination to reverse multidrug resistance in leukemia.
ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; biosynthesis ; ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; Antineoplastic Agents ; pharmacology ; Benzylisoquinolines ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Humans ; K562 Cells ; Leukemia ; metabolism ; pathology ; Multidrug Resistance-Associated Proteins ; biosynthesis ; Neoplasm Proteins ; biosynthesis ; Protein Array Analysis ; Tamoxifen ; analogs & derivatives ; pharmacology

BACKGROUNDNon-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.

METHODSOne 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.

RESULTSTransmission 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).

CONCLUSIONSThere 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.

Adult ; Aged ; Blood Platelets ; ultrastructure ; Carcinoma, Non-Small-Cell Lung ; blood ; drug therapy ; mortality ; ultrastructure ; Female ; Humans ; Male ; Microscopy, Electron, Transmission ; Middle Aged ; P-Selectin ; blood ; Survival Analysis

Objective To explore the relationships between the polyrnorphisms of xeroderma pigmentosum A(XPA) and the susceptibility of esophageal cancer (EC),as well as its interaction with environmental factors-gene in Changzhi area,Shanxi province. Methods A case-control study was conducted,including 196 cases of EC and 201 controls.XPA 23G polymorphisms were determined with polymerase chain-restriction on fragment length polymorphism (PCR-RFLP).Results The risk of EC was significantly degraded in the individuals who had been carrying the XPA heterozygote (A/G) and mutation genotype (G/G),compared to those with wild genotype (X2=16.21,P＜0.01) and the ORs were 0.58 (0.37-0.91) and 0.32 (0.18-0.56),respectively.There was negative interaction between XPA 23G mutation genotype and the consumption of pickled food (S=0.04,API=-0.77).Conclusion Genetic polymorphism in the XPA 23G might be associated with esophageal cancer in Changzhi area,and there was a negative action between XPA predisposing genotype and the consumption of pielded food.

After the epidemic of novel Coronavirus Disease 2019(COVID-19), construction of disease prevention and control has become a top priority. As a pioneer in the recovery of global economy and society, Shanghai should play a fundamental role in building a comprehensive system of public health and advanced disease prevention and control in the new era. In this article, we systematically categorize the requirements for the construction of disease prevention and control system in the new era, identify the weakness and challenges during and after the epidemic, and then make suggestions. It is proposed that we should utilize the important window period of the"14th Five-Year Plan", with the"Healthy China"strategy and municipal"20 Tasks for Public Health Construction"as the starting point, to make substantial contribution to the functional orientation, investment of resources, capacity building, operational mechanism and team building, which may provide scientific evidence for the reform and development of disease prevention and control system.

OBJECTIVETo evaluate the feasibility and practicality of laparoscopic hepatectomy for hepatic hemangioma.

METHODSCandidate for laparoscopic liver resection were 18 cases of hepatic hemangioma from January 2002 to October 2006. The portal bloods stream was blocked by the laparoscope portal blood blocker. The Electric-cautery and ultracision were used for liver transection. Operative procedures included anatomical left hepatectomy in 2 cases, non-anatomical left hepatectomy 1 case, left lobectomy 5 cases, local liver resection 10 cases. Two cases of hepatic hemangioma associated with gallbladder stone were performed cholecystectomy synchronously, 1 case associated with chronic appendicitis were performed appendectomy synchronously.

RESULTSLaparoscopic left liver resection was successfully performed in all 18 cases. The operative duration was (185.4 +/- 55.7) min. The quantity of blood lost during the operation was (416.2 +/- 128.8) ml. The postoperative recovery was smooth and good. No critical complications occurred. The duration for hospitalization was (6.2 +/- 1.0) d.

CONCLUSIONLaparoscope hepatectomy for hepatic hemangioma is safe and feasible.

Adult ; Feasibility Studies ; Female ; Hemangioma ; surgery ; Hepatectomy ; methods ; Humans ; Laparoscopy ; Liver Neoplasms ; surgery ; Male ; Middle Aged ; Treatment Outcome

The aim of this study was to set up a new method for 5, 10-Methylenetetrahydrofolate reductase (MTHFR) single nucleotide polymorphisms (SNP) genotyping, and to investigate the hereditary susceptibility of hematological malignancy. Prepared an aimed gene microarray based on cDNA microarray theory, dual-color fluorescence hybridization was used to detect SNP loci, and DNA sequencing was performed to confirm the results. The MTHFR C677T SNP loci of 157 controls and 127 patients with hematological malignancies (30 multiple myeloma, 28 non-Hodgkin's lymphoma, 22 acute lymphoblastic leukemia, 40 acute myeloid leukemia, 7 chronic myeloid leukemia) from Jiangsu province were detected. The results showed that after overlapping, homozygous wild type, heterozygote type and homozygous mutant type yielded green, yellow and red fluorescence, respectively. DNA sequencing validated these results. The allele frequency of 677C and 677T in patients and controls were 58.7% and 66.9%, 41.3% and 33.1% respectively, showing statistically significant difference (chi2 = 4.077, P = 0.043). 677TT genotype showed a significantly higher risk of MM (OR = 4.21; 95% CI = 1.50 - 11.83; P = 0.006). It is concluded that this microarray-based method is accurate, high-throughput and inexpensive, suitable for SNP genotyping in a large number of individuals. C677T polymorphisms influence the risk of hematological malignancies. 677TT genotype is susceptive to MM.
Adult ; Aged ; Base Sequence ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Hematologic Neoplasms ; enzymology ; genetics ; Humans ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Middle Aged ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Polymorphism, Single Nucleotide

OBJECTIVETo explore the synthetical index for diagnosing schistosomiasis with ultrasound and to assess the prevalence rate with the index.

METHODSUltrasound indexes of schistosomiasis Japonicum were analyzed by principal component analysis, and the synthetical indexes were assessed by ROC curve.

RESULTSAmong the abnormal rates of the 6 indexes, the lowest was 1.6% comparing with the highest of 59.5%. Significant difference was noficed among the abnormal rates (chi(2) = 631.1, P < 0.01). The individual correlation of the six indexes to each other as will as with age distribution was significant (P < 0.05). The three principal components reflected the degree of pathological changes on liver and spleen. The first principal component was the factor reflecting the degree of liver pathological changes, and the second and third principal components reflected the degree of pathological changes on spleen. The synthetical index D(1) = 0.047X(1) + 0.428X(2) + 1.247X(3) + 0.095X(4) + 0.002X(5) + 0.213X(6) - 12.837 was found by adding the three weight principal components, and it's area under the ROC curve was 0.957. When -1.70 was taken as the critical value, the abnormal rate of population was 66.3%, close to the resident's actual prevalence rate 66.9%.

CONCLUSIONUltrasonography was considered as a method which could rapidly assessing the resident's prevalence rate in the endemic areas of schisitosomiasis Japonicum, and could also provide powerful information for development of strategy on chemotherapy.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Principal Component Analysis ; ROC Curve ; Schistosomiasis japonica ; diagnostic imaging ; epidemiology ; Ultrasonography