Objective: To explore the effects of exogenous APMCF1 gene on the growth and cell cycle of HHCC cells. Methods: A full human APMCF1 gene was cloned into expression vector pcDNA3.0 and transfected into human hepatocellular carcinoma cell HHCC by lipofectamine,and the positive clone was screened by G418.The expression of APMCF1 protein in HHCC cells were examined by semi-quantitative RT-PCR,and its effects on cell growth and cell cycle were observed by MTT and flow cytometer(FCM). Results: Semi-quantitative RT-PCR indicated that APMCF1 had been successfully transfected into HHCC cells.The growth speed of APMCF1 infected cells decreased markedly.Analysis of cell cycle by FCM showed that 85.3% of the infected cells were in G_1 phase and 12.5% in S phase,while 66.4% of the control HHCC cells were in G_1 phase and 24.7% in S phase. Conclusion:Exogenous APMCF1 gene can inhibit the growth and cell cycle of human hepatocellular carcinoma cell HHCC.It may be a cell cycle regulative gene.

AIM: To determine the relationship between the mutation of the RII gene and RER status in the tumorigenesis of sporadic colorectal cancer. METHODS: We screened RER status and mutation of the RII gene from 50 sporadic colorectal cancers (19 in the proximal colon, 31 in the distal colorectum). RESULTS: RER was found in 13 cases (8 in the proximal colon, 5 in the distal colorectum), and 5 of them showed mutations of the RII gene. All 5 cancers carrying a TGF-? RII gene mutation showed RER+, but there wasn't any mutation of RII gene in RER(-) cases. Four of 5 RII mutation were located at the cecum. CONCLUSION: These data indicate that the TGF-? RII gene is a major target of microsatellite instability and mutation of the RII gene play an important role in carcinogenesis of sporadic colorectal cancer with microsatellite instability, especially at the cecum. [

Objective To investigate the role and the mechanism of Apr-1 gene on cholangiocarcinoma QBC939 cell lines proliferation and cell cycle regulation. Methods Apr-1 gene was transfected into QBC939 cells by using liposomes to establish a QBC939 cell model ( QBC939-Apr-1 ) stably expressing Apr-1 gene. Apr-1 mRNA expression and the changes in cell cycle and cell growth of QBC939 cells were analyzed by RT-PCR, flow cytometry ( FCM ) and growth curve before and after transfection. The regulatory effect of Apr-1 gene on the expression of cell cycle-related genes was investigated in QBC939 cells before and after Apr-1 transfection using cell cycle gene microarrays. Results Significant suppression of cell growth was observed with the cell model stably expressing Apr-1 gene. Apr-1 over-expression caused cell arrest from 9% to 13% (P ＜0. 01 ) increase in G2 population. Cell cycle gene microarrays demonstrated that the expression of Skp2 、UBE1 was up-regulated, while the expression of MRE11A 、CKS2 、CDK8 、CDC45 was down-regulated by more than 3 folds. Conclusions Apr-1 gene suppresses QBC939 cell proliferation in vitro, QBC939 cells presented with differences in the expression of cell cycle-related genes after Apr-1 gene transfection.

Leaf senescence is considered as one of important factors to decrease ornamental values of foliage plants. In the attempt to study and understand the molecular mechanism of leaf senescence, a senescent leaf cDNA library of Coleus blumei was constructed and a small EST library was obtained. According to the sequence of an EST fragment with a cystathionine beta synthase (CBS) domain, a novel leaf senescenceassociated gene (SAG) full-length cDNA encoding a CBS-domain-containing protein, denoted Cbcbs, was rapidly cloned using a strategy of RACE combined with cDNA library. The full length of the Cbcbs gene was 859 bp long (accession No. EF076754) and contained a 609 bp open reading frame (ORF) encoding a 202amino acid protein. One stop codon (TAA) was found in 5' UTR and one possible polyadenylation signal,AATAAA, and a pentanucleotide motif, ATTTA, were found in 3' UTR. The CbCBS contained a predicted mitochondrial targeting peptide in the N-terminal region, two conserved and intact CBS domains, four casein kinase Ⅱ (CK Ⅱ) phosphorylation sites, three protein kinase c (PKC) phosphorylation sites and one tyrosine sulfation (TS) site. Sequence comparisons and phylogenetic analysis showed that CbCBS was a novel senescence or stress-associated protein. The prediction analysis of secondary structure and three dimensional structure of CbCBS suggested that the chief function of the protein was decided by the CBS domain pair. The expression pattern of Cbcbs in leaves was analyzed by RT-PCR. It was demonstrated that Cbcbs gene was a senescence-associated gene (SAG) and expressed in all leaf stages, young stage (Y) being the lowest and terminal senescence stage (S3) being the highest, and was upregulated along with the leaf senescence.Function analysis showed that the mature CbCBS maybe acts as a sensor of cellular energy status and directly or indirectly regulates cellular energy levels to increase ATP content in mitochondria during periods of metabolic stress of senescent leaves.

One hundred and ten patients with primary hepatocellular carcinoma underwent hepatectomy from 2005 to 2010.Different methods of hepatic exclusion were applied in the surgery,including 54 patients with total hepatic vascular exclusion,22 with half hepatic vascular exclusion and 34 with regional hepatic vascular exclusion.The recovery of postoperative liver function was retrospectively analyzed.The results showed that the postoperative liver function of regional hepatic vascular exclusion [ALT(311 ± 80) U/L,total bilirubin (TB) (22.2 ± 8.3) μmoL/L at 1 st day and ALT (58 ± 17) U/L,TB (11.3 ± 3.1)μmol/L at 7th day] was better than that of total hepatic vascular exclusion [ALT(874 ±299)U/L,TB (42.9± 19.1) μmol/L at 1st day (P＜0.05) and ALT (108-±52)U/L,TB (14.6±9.2) μmol/L at 7th day] (P ＜ 0.05,＞ 0.05),indicating that regional hepatic vascular exclusion can effectively reduce hepatic injury during the operation and promote recovery of liver function after operation.

Objective:To analyze the epidemiological characteristics of novel coronavirus positive cases including confirmed cases with clinical symptoms and asymptomatic infected cases in Guangzhou.Methods:Epidemiological data were collected on the nucleic acid positive cases of COVID-19 in Guangzhou from January to September 2020. The epidemiological characteristics, the distribution of time intervals between the confirmed/isolation date and the date of the first positive detection were analyzed, at last the influencing factors for the confirmed cases and asymptomatic infected persons were discussed.Results:From January 7 to September 4 in 2020, a total of 1 097 nucleic acid positive cases were identified, including 658 confirmed cases (59.98%) and 439 asymptomatic infected cases (40.02%). Among the 658 confirmed cases, the median age was 42 years old, the cases indicated two significant peaks. one of the peaks was related to the imported and associated cases from Hubei province, and the other peak was connected with individuals from overseas. In terms of 439 asymptomatic infected cases, the median age was 32 years old. There were two stages in these cases. The first stage followed the second peak of confirmed cases, and the second stage overlapped with the confirmed cases in Guangzhou when the epidemic was in a period of normal prevention and control, mainly related to imported cases from abroad. The asymptomatic infected persons accounted for 57.32% in all the imported infected cases. In both of asymptomatic and symptomatic cases, the positive rate of pharyngeal swabs was higher than that of nasopharyngeal swabs and anal swabs. There were statistically significant differences in age, source of infection and gender composition between confirmed cases and asymptomatic infected persons ( P<0.05). Older age groups were more likely to have clinical symptoms, with ≥40 years being the risk factor for confirmed cases (OR=2.334, P=0.001), and 20-39 years less likely to have clinical symptoms (OR=0.620, P=0.047), compared with the 0-19 years old group. Compared with those infected in China, those infected abroad were less likely to develop clinical symptoms and became confirmed cases (OR=0.723, P=0.013). Women were more likely to have clinical symptoms than men (OR=1.574, P=0.001). Conclusions:At present, asymptomatic infected persons and confirmed patients with clinical symptoms co-existed, and the number of asymptomatic infected patients was higher than that of confirmed cases in Guangzhou. High age, domestic infection and female may be risk factors for confirmed cases. It was of great value to further explore these underlying mechanisms for the prevention and treatment of the COVID-19.

Objective To evaluate the efficacy of a sustained releasing mosquito larvicide package against larval breeding and its impact on water and plant,in order to provide a scientific evidence for its application in control and prevention of Dengue.Methods Guangzhou Center for Disease Control and Prevention was chosen as the test place.Twenty test sites were set up,2 bags of sustained releasing larvicides package,1 bag of sustained releasing larvicides package,3 g 1％ temephos granules and nothing were put into 4 glass bottles for each test site from July to December in 2014,respectively.The 4 glass bottles were called high dose (H) group,low dose (L)group,positive control (P) group and blank control (B) group,respectively.The 4 groups were observed at intervals of 10 days for 19 times.Environmental air temperature,turbidity of water,number of larvae and damage of plant were recorded.And 5 test sites were selected to collect water specimen.The chemical oxygen demand,ammonia nitrogen concentration and temephos concentration of water specimen were detected.Results The larval breeding rates were 0 (0/380),1.1％ (4/380),0.8％ (3/380) and 63.4％ (241/380),damage rates of plant were 5.0％ (19/380),5.5％ (21/380),4.7％ (18/380),4.7％ (18/380) and turbidty rates of water were 24.5％ (93/380),19.7％ (75/380),33.4％ (127/380) and 20.3％ (77/380) in H,L,P and B groups,respectively.Statistically significant differences were seen in larval breeding rate and turbidity rate of water between different groups (x2 =823.565,24.715,all P ＜ 0.05),but they were not seen in damage rate of plant (x2 =0.332,P ＞ 0.05).The temephos concentrations were 1.24,0.78 and 2.33 mg/L in H,L and P groups,respectively.Statistically significant differences were seen in temephos concentration between different groups (H =35.426,P ＜ 0.01),but they were not seen in chemical oxygen demand and ammonia nitrogen concentration (H =0.239,0.013,all P ＞ 0.05).Conclusions The sustained releasing package of mosquito larvicide makes less pollution to water and has no impact on water turbidity.Moreover,it doesn't damage the aquatic plant.The efficacy of the sustained releasing package of mosquito larvicide could effectively prevent mosquito larval breeding in Dengue epidemic period.

Using RACE with a Fagopyrum dibotrys callus cDNA library, one clone, named FdMYBP1, encoding a putative R2R3 MYB protein was identified. FdMYBP1 appeared to be a full-length cDNA of 1159 bp encoding a protein of 265 amino acids. Through structure and property analysis of FdMYBPI with bioinformational methods, it was found that the amino acid sequence of FdMYBP1 showed great homology to other MYBP with the R2R3 repeat region in the N-terminus. Southern blot analysis indicated that FdMYBP1 belongs to a single copy gene in F. dibotrys genomes. The FdMYBP1 gene has the same classic characters with other MYBP and probably involved in the pathway of flavonoid metabolisms.
Cloning, Molecular ; Fagopyrum ; genetics ; metabolism ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Plant Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-myb ; genetics ; metabolism

The fruit fly, Drosophila melanogaster, is able to discriminate visual landmarks and form visual long-term memory in a flight simulator. Studies focused on the molecular mechanism of long-term memory have shown that memory formation requires mRNA transcription and protein synthesis. However, little is known about the molecular mechanisms underlying the visual learning paradigm. The present study demonstrated that both spaced training procedure (STP) and consecutive training procedure (CTP) would induce long-term memory at 12 hour after training, and STP caused significantly higher 12-h memory scores compared with CTP. Label-free quantification of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and microarray were utilized to analyze proteomic and transcriptomic differences between the STP and CTP groups. Proteomic analysis revealed 30 up-regulated and 27 down-regulated proteins; Transcriptomic analysis revealed 145 up-regulated and 129 down-regulated genes. Among them, five candidate genes were verified by quantitative PCR, which revealed results similar to microarray. These results provide insight into the molecular components influencing visual long-term memory and facilitate further studies on the roles of identified genes in memory formation.
Animals ; Conditioning (Psychology) ; physiology ; Drosophila Proteins ; genetics ; metabolism ; Drosophila melanogaster ; genetics ; metabolism ; physiology ; Flight, Animal ; physiology ; Gene Expression Profiling ; methods ; Memory ; physiology ; Oligonucleotide Array Sequence Analysis ; Proteomics ; methods ; Time Factors ; Vision, Ocular ; physiology

OBJECTIVETo establish a method for isolation of the total RNA from Fagopyrum cymosum callus.

METHODThe improved method combining that of CTAB extraction with the LiCl precipitation was used to isolate the total RNA from the four F. cymosum callus. The quality of the RNA was detected by UV spectrophotometric analysis, 0.8% non-denaturing agarose gel electrophoresis and RNA reverse transcription.

RESULTThe bands of 28S and 18S could be seen clearly by agarose gel electrophoresis, and the value of A260/A280 was between 1.9 and 2.0. The cDNA which was reverse-transcribed by the total RNA showed a wide length rage of 500 bp-5 kb.

CONCLUSIONThe RNA extracted by this method meets the requirement of reverse transcription-polymerase chain reaction (RT-PCR), construction of cDNA libraries, et al. This improved method can be used to isolate the total RNA from F. cymosum callus with the advantage of simpleness, efficiency and low cost.