Leptin-mediated ERK Signaling Pathway Promotes the Transformation of Rat Alveolar Type II Epithelial Cells Induced by Yunnan Tin Mine Dust / 中国肺癌杂志

BACKGROUND@#Currently, a significant number of miners are involved in mining operations at the Gejiu tin mine in Yunnan. This occupational setting is associated with exposure to dust particles, heavy metals, polycyclic aromatic hydrocarbons, and radioactive radon, thereby significantly elevating the risk of lung cancer. This study aims to investigate the involvement of leptin-mediated extracellular regulated protein kinase (ERK) signaling pathway in the malignant transformation of rat alveolar type II epithelial cells induced by Yunnan tin mine dust.@*METHODS@#Immortalized rat alveolar cells type II (RLE-6TN) cells were infected with Yunnan tin mine dust at a concentration of 200 μg/mL for nine consecutive generations to establish the infected cell model, which was named R₂₀₀ cells. The cells were cultured normally, named as R cells. The expression of leptin receptor in both cell groups was detected using the Western blot method. The optimal concentration of leptin and mitogen-activated protein kinase kinase (MEK) inhibitor (U0126) on R₂₀₀ cells was determined using the MTT method. Starting from the 20th generation, the cells in the R group were co-cultured with leptin, while the cells in the R₂₀₀ group were co-cultured with the MEK inhibitor U0126. The morphological alterations of the cells in each group were visualized utilizing hematoxylin-eosin staining. Additionally, concanavalin A (ConA) was utilized to detect any morphological differences, and an anchorage-independent growth assay was conducted to assess the malignant transformation of the cells. The changes in the ERK signaling pathway in epithelial cells after the action of leptin were detected using the Western blot method.@*RESULTS@#Both the cells in the R group and R₂₀₀ group express leptin receptor OB-R. Compared to the R₂₀₀ group, the concentration of leptin at 100 ng/mL shows the most significant pro-proliferation effect. The proliferation of R₂₀₀ cells infected with the virus is inhibited by 30 μmol/L U0126, and a statistically significant divergence was seen when compared to the control group (P<0.05). Starting from the 25th generation, the cell morphology of the leptin-induced R₂₀₀ group (R₂₀₀L group) underwent changes, leading to malignant transformation observed at the 30th generation. The characteristics of malignant transformation became evident by the 40th generation in the R₂₀₀L group. In contrast, the other groups showed agglutination of P40 cells, and the speed of cell aggregation increased with an increase in ConA concentration. Notably, the R₂₀₀L group exhibited faster cell aggregation compared to the U0126-induced R₂₀₀ (R₂₀₀LU) group. Additionally, the cells in the R₂₀₀L group were capable of forming clones starting from P30, with a colony formation rate of 2.25‰±0.5‰. However, no clonal colonies were observed in the R₂₀₀LU group and R₂₀₀ group. The expression of phosphorylated extracellular signal-regulated kinase (pERK) was enhanced in cells of the R₂₀₀L group. However, when the cells in the R₂₀₀L group were treated with U0126, a blocking agent, the phosphorylation level of pERK decreased.@*CONCLUSIONS@#Leptin can promote the malignant transformation of lung epithelial cells infected by mine dust, and the ERK signaling pathway may be necessary for the transformation of alveolar type II epithelial cells induced by Yunnan tin mine dust.

Study on the role of CCL19 and AKT signaling pathway in the development of lung cancer / 中国职业医学

Objective To investigate the role of chemokine ligand 19 (CCL19) and protein kinase-B (AKT) signaling pathway in lung cancer development. Methods The human lung adenocarcinoma cell line, A549 cells, in logarithmic growth phase were randomly divided into five groups： blank control group, solvent control group, CCL19 treatment group, AKT inhibition group, and antibody neutralization group. The blank control group received no treatment. The other four groups were treated with dimethyl sulfoxide, CCL19, MK-2206 (AKT inhibitor), and a combination of CCL19 and MK-2206, respectively. Cell viability was assessed using the CCK-8 assay, while cell migration and invasion capabilities were evaluated using the cell scratch and transwell assays. The relative expression levels of Pan-AKT, p-AKT (Ser473), p-AKT (Thr308), E-cadherin (E-cad), N-cadherin (N-cad), and Snail proteins in A549 cells were detected using Western blotting. Lung cancer tissue samples from 60 patients with non-small cell lung cancer (NSCLC) were collected, and the expression of CCL19 and matrix metalloproteinase 9 (MMP9) proteins in the specimens was examined using immunohistochemistry. Results The survival rate of A549 cells in the AKT inhibition group and antibody neutralization group was lower than that in blank control group, solvent control group, and CCL19 treatment group (all P<0.05). The cell scratch assay result showed that the cell migration rate of the CCL19 treatment group was higher at 36.0 and 48.0 hours than those of the blank control group, solvent control group, AKT inhibition group, and neutralizing antibody group (all P<0.05). The Transwell assay result showed that the invasion amount of A549 cells in the AKT inhibition group was less than that in the CCL19 treatment group (P<0.05). Compared with the blank control group, the relative expression of E-cad protein in the CCL19 treatment group decreased, while the relative expression of p-AKT (Ser473), p-AKT (Thr308), N-cad and Snail proteins increased (all P<0.05). The relative expression of p-AKT (Ser473), p-AKT (Thr308), N-cad, and Snail proteins in A549 cells decreased (all P<0.05), and relative expression of E-cad protein increased (all P<0.05) in the AKT inhibition group and antibody neutralization group compared with the blank control group, solvent control group, and CCL19 treatment group. There was no significant difference in the expression of CCL19 and MMP9 in lung cancer tissues of NSCLC patients in Xuanwei City, Gejiu City, and other regions (all P>0.05). The expression of CCL19 and MMP9 in NSCLC patients with lymph node metastasis was higher than in patients without lymph node metastasis (all P<0.01). Conclusion CCL19 can promote the invasion and metastasis of lung cancer cells and induce epithelial-mesenchymal transition. Its expression level is related to lymph node metastasis in NSCLC patients. The AKT signaling pathway may be an important mechanism underlying lung cancer development.

Regulation of LncRNA00067110 on Proliferation, Apoptosis and Melanins Production of B16-F10 Cells by Targeting Cabyr / 中国生物化学与分子生物学报

Long non-coding RNA (lncRNA) is a type of non-coding RNA with the more than 200 nucleotides. Several lncRNAs have been identified as the potential targets for cancer therapy. LncRNA00067110 is one of the differentially expressed genes in the transcriptome profiles of melanoma B16-F10 cells compared to normal mice melanocytes. To investigate whether lncRNA00067110 regulates the proliferation, apoptosis and melanogenesis of B16-F10 cells, the calcium-binding tyrosine phosphorylation regulated protein (Cabyr) target gene was predicted by LncTar and verified by dual luciferase activities. The regulating function of lncRNA00067110 was investigated by the analysis of transcriptome profiles and to detect the proliferation, apoptosis and melanin production of B16-F10 cells transfected by the overexpression plasmids of lncRNA00067110. The results showed that the relationship of lncRNA00067110 targeting Cabyr, the mRNA and protein levels of proliferation (MEK/ERK/MNK/CREB) and melanogenesis-related genes (TYR family and CREB) were significantly down-regulated, while the mRNA and protein levels of apoptosis-related genes (AKT and Bcl-2) were up-regulated in B16-F10 cells with lncRNA00067110 overexpression. The transcriptome profile of B16-F10 cells with lncRNA00067110 overexpression showed that 17 genes were differentially expressed, among which Cabyr was up-regulated. Furthermore, the effect of lncRNA00067110 on the phenotypes of cell proliferation and apoptosis were verified. The results suggested that lncRNA00067110 might be a novel target for the treatment of melanoma by targeting Cabyr, which regulate the expression of related genes to inhibit the proliferation and melanogenesis, as well as to induce the apoptosis of B16-F10 cells.

Epidemiological characteristics of local COVID-19 epidemics and control experience in routine prevention and control phase in China / 中华流行病学杂志

The COVID-19 pandemic is still ongoing in the world, the risk of COVID-19 spread from other countries or in the country will exist for a long term in China. In the routine prevention and control phase, a number of local COVID-19 epidemics have occurred in China, most COVID-19 cases were sporadic ones, but a few case clusters or outbreaks were reported. Winter and spring were the seasons with high incidences of the epidemics; border and port cities had higher risk for outbreaks. Active surveillance in key populations was an effective way for the early detection of the epidemics. Through a series of comprehensive prevention and control measures, including mass nucleic acid screening, close contact tracing and isolation, classified management of areas and groups at risk, wider social distancing and strict travel management, the local COVID-19 epidemics have been quickly and effectively controlled. The experiences obtained in the control of the local epidemics would benefit the routine prevention and control of COVID-19 in China. The occurrence of a series of COVID-19 case clusters or outbreaks has revealed the weakness or deficiencies in the COVID-19 prevention and control in China, so this paper suggests some measures for the improvement of the future prevention and control of COVID-19.

Course of disease and related epidemiological parameters of COVID-19: a prospective study based on contact tracing cohort / 中华预防医学杂志

Objective: To analyze the course of disease and epidemiological parameters of COVID-19 and provide evidence for making prevention and control strategies. Methods: To display the distribution of course of disease of the infectors who had close contacts with COVID-19 cases from January 1 to March 15, 2020 in Guangdong Provincial, the models of Lognormal, Weibull and gamma distribution were applied. A descriptive analysis was conducted on the basic characteristics and epidemiological parameters of course of disease. Results: In total, 515 of 11 580 close contacts were infected, with an attack rate about 4.4%, including 449 confirmed cases and 66 asymptomatic cases. Lognormal distribution was fitting best for latent period, incubation period, pre-symptomatic infection period of confirmed cases and infection period of asymptomatic cases; Gamma distribution was fitting best for infectious period and clinical symptom period of confirmed cases; Weibull distribution was fitting best for latent period of asymptomatic cases. The latent period, incubation period, pre-symptomatic infection period, infectious period and clinical symptoms period of confirmed cases were 4.50 (95%CI:3.86-5.13) days, 5.12 (95%CI:4.63-5.62) days, 0.87 (95%CI:0.67-1.07) days, 11.89 (95%CI:9.81-13.98) days and 22.00 (95%CI:21.24-22.77) days, respectively. The latent period and infectious period of asymptomatic cases were 8.88 (95%CI:6.89-10.86) days and 6.18 (95%CI:1.89-10.47) days, respectively. Conclusion: The estimated course of COVID-19 and related epidemiological parameters are similar to the existing data.

Epidemiological characteristics and spatial epidemiology of hand-foot-mouth disease in Hunan Province, China, from 2008 to 2019 / 中国当代儿科杂志

OBJECTIVES@#To investigate the epidemic situation of hand-foot-mouth disease (HFMD) in Hunan Province, China, from 2008 to 2019, as well as its spatial autocorrelation characteristics and spatial-temporal clustering, and to provide a reference for the prevention and control of HFMD in Hunan Province.@*METHODS@#Spatial autocorrelation and spatial-temporal clustering analyses were used to analyze the monitoring data of HFMD in Hunan Province from 2008 to 2019.@*RESULTS@#The epidemic situation of HFMD in Hunan Province from 2008 to 2019 showed obvious seasonal distribution, with a low incidence rate in January to March and a high incidence rate in April to July. As for population distribution, children aged 0-5 years had the highest number of HFMD cases and accounted for 95.89% (1 460 391/1 522 910) of all cases, with a mean annual incidence rate of 2 197.784/100 000, and scattered children had the highest number of cases and accounted for 82.59% (1 257 739/1 522 910) of all cases. The global spatial autocorrelation analysis showed that the onset of HFMD in Hunan Province showed a significant clustering distribution, and the local spatial autocorrelation analysis showed that the high clustering areas of HFMD were mainly the districts and counties of Changsha, Zhuzhou, and Yueyang cities. Time-space scanning showed that clustering time was mainly April to July; the cases were clustered in the northeast of Hunan Province from 2008 to 2010 and in the central part of Hunan Province from 2011 to 2019.@*CONCLUSIONS@#The high incidence rate of HFMD is observed in April to July in Hunan Province. Children under 5 years of age are at a high risk of this disease. Spatial-temporal clustering is observed for the epidemic of HFMD, mainly clustered in the northeastern and central areas of Hunan Province. It is suggested that the results may be used as guidance to determine the key areas for HFMD prevention and control in Hunan Province and optimize the allocation of health resources.

MicroRNA-100-5p Regulates Melanogenesis in Alpaca Melanocytes by Targeting FGF21 / 中国生物化学与分子生物学报

Although many microRNAs (miRNAs) are known to function as regulators of coat color and melanogenesis, the underlying molecular mechanisms of miR-100-5p governing melanogenesis were not completely known.The goal of this study was to determine the effect of miR-l()()-5p on melanogenesis in alpaca melanocytes.Fibroblast growth factor 21 (FGF21) is a predicted target gene of miR-100-5p and the luciferase reporter assay demonstrated that miR-100-5p regulates FGF21 by binding to its 3' untranslated region (3'UTR).In this study, alpaca melanocytes were transfected with miR-100-5p, inhibitor and negative control plasmid.Results showed that miR-100-5p overexpression significantly decreased mRNA and protein expression of FGF2\.Meanwhile, the ERK signal pathway was inhibited, with subsequent up-regulation of microphthalmia-associated transcription factor (MITF) , tyrosinase (TYR) and tyrosinase-related protein 2 (TYRP2), which increased melanin production.The results suggest that miR-100-5p may regulate melanogenesis by targeting FGF21 via extracellular regulated MAP kinase (ERK) signaling pathway.

DKK3 Regulates Melanogenesis in Melanocytes of Alpaca / 中国生物化学与分子生物学报

Dickkopf-3 (DKK3) , as a critical inhibitor of the Wnt/p-catenin signaling pathway, may he involved in melanogenesis.In the current study, we investigated the effects of DKK3 on melanogenesis in melanocytes of alpaca.Overexpression of DKK3 in alpaca melanocytes, the expression of Wntl, Lefl , Myc and the major target genes termed microphthalmia-associated transcription factor (M1TF) and its downstream genes, including tyrosinase (TYR), tyrosinase-related protein 1 (TYRP1) and tyrosinase- related protein 2 (TYRP2) were significantly decreased at both mRNA and protein levels (P<0.05); total alkali melanin, pheomelanin and eumelanin were decreased by 80.30%, 72.17% and 64.60% (P <0.05), respectively.In contrast, in the melanocytes transfected with siRNA-DKK3 (a small interference RNA targeting DKK3) , the expression of Wntl, Lefl, Myc, MITF, TYR, TYRPl and TYRP2 were significantly increased at both mRNA and protein levels (P<0.05) ; total alkali melanin, pheomelanin and eumelanin were significantly increased by 1.65 folds, 1.25 folds and 1.21 folds (P< 0.05) , respectively.These results indicate that DKK3 regulates melanogenesis in alpaca melanocytes via the Wnt/p-catenin signaling pathway and down-regulates MITF.

TMEM106B Induces Melanogenesis by Regulating ERK/CREB Signaling Pathway / 中国生物化学与分子生物学报

The TMEM106B protein is a type-II transmembrane protein, which localizes in the endosome and lysosome of dendrites in primary neurons. TMEM106B is essential for maintaining and branching of dendrites, and thus regulates retrograde lysosomal trafficking of dendrites in primary neurons. Mammalian melanocytes are derived from neural cells, while melanosomes are originated from early endosome. However, the function of TMEM106B protein in melanocytes and its potential molecular mechanism in melanogenesis still remain unknown. Recently it was reported that transcription factor EB (TFEB) was the regulator of lysosome synthesis and TMEM106B protein overexpression promoted TFEB translocation into the nucleus. However, MITF (microphthalmia-associated transcription factor) and TFEB regulate each other in melanoma cells in vitro. Here in, plasmid containing gene for TMEM106B overexpression was transfected into melanocytes to investigate the regulation of TMEM106B on melanogenesis. The results showed that TMEM106B protein was localized in the cytoplasm of melanocytes. Compared with the negative control (NC), the mRNA levels of cyclic AMP-responsive element-binding protein (CREB) and MITF, especially CREB, were significantly increased in melanocytes with TMEM106B overexpression P< 0. 001). Western blot analysis showed that the expression of phosphorylated MAP kinase (p-ERK) was apparently increased (P<0.001) and resulted in the up-regulation of melanogenic regulatory proteins, including MITF, tyrosinase (TYR), tyrosinase-related protein-1 (TYRP1) and 2 (TYRP2). Masson-Fontana method showed that TMEM106B influenced the production of melanin in melanocytes. The spectrophotometry assay indicated that the amount of total melanin (ASM) (P<0. 001) and eumelanin (EM) (P<0. 05) were increased in alpaca melanocytes transfected with TMEM106B, while pheomelanin (PM) (P<0. 001) was decreased. These results demonstrated that TMEM106B played a vital role in melanogenesis in melanocytes by regulating ERK/CREB signaling pathway.

Cross-neutralizing Anti-hemagglutinin Antibodies Isolated from Patients Infected with Avian Influenza A (H5N1) Virus / 生物医学与环境科学（英文）

Objective@#To recover broad-neutralizing monoclonal antibodies (BnAbs) from avian influenza A (H5N1) virus infection cases and investigate their genetic and functional features.@*Methods@#We screened the Abs repertoires of expanded B cells circulating in the peripheral blood of H5N1 patients. The genetic basis, biological functions, and epitopes of the obtained BnAbs were assessed and modeled.@*Results@#Two BnAbs, 2-12D5, and 3-37G7.1, were respectively obtained from two human H5N1 cases on days 12 and 21 after disease onset. Both Abs demonstrated cross-neutralizing and Ab-dependent cellular cytotoxicity (ADCC) activity. Albeit derived from distinct Ab lineages, , V 1-69-D2-15-J 4 (2-12D5) and V 1-2-D3-9-J 5 (3-32G7.1), the BnAbs were directed toward CR6261-like epitopes in the HA stem, and HA I45 in the hydrophobic pocket was the critical residue for their binding. Signature motifs for binding with the HA stem, namely, IFY in V 1-69-encoded Abs and LXYFXW in D3-9-encoded Abs, were also observed in 2-12D5 and 3-32G7.1, respectively.@*Conclusions@#Cross-reactive B cells of different germline origins could be activated and re-circulated by avian influenza virus. The HA stem epitopes targeted by the BnAbs, and the two Ab-encoding genes usage implied the VH1-69 and D3-9 are the ideal candidates triggered by influenza virus for vaccine development.

Risk assessment of exported risk of novel coronavirus pneumonia from Hubei Province / 中华预防医学杂志

To evaluate the exported risk of novel coronavirus pneumonia (NCP) from Hubei Province and the imported risk in various provinces across China. Data of reported NCP cases and Baidu Migration Indexin all provinces of the country as of February 14, 2020 were collected. The correlation analysis between cumulative number of reported cases and the migration index from Hubei was performed, and the imported risks from Hubei to different provinces across China were further evaluated. A total of 49 970 confirmed cases were reported nationwide, of which 37 884 were in Hubei Province. The average daily migration index from Hubei to other provinces was 312.09, Wuhan and other cities in Hubei were 117.95 and 194.16, respectively. The cumulative NCP cases of provinces was positively correlated with the migration index derived from Hubei province, also in Wuhan and other cities in Hubei, with correlation coefficients of 0.84, 0.84, and 0.81. In linear model, population migration from Hubei Province, Wuhan and other cities in Hubei account for 71.2%, 70.1%, and 66.3% of the variation, respectively. The period of high exported risk from Hubei occurred before January 27, of which the risks before January 23 mainly came from Wuhan, and then mainly from other cities in Hubei. Hunan Province, Henan Province and Guangdong Province ranked the top three in terms of cumulative imported risk (the cumulative risk indices were 58.61, 54.75 and 49.62 respectively). The epidemic in each province was mainly caused by the importation of Hubei Province. Taking measures such as restricting the migration of population in Hubei Province and strengthening quarantine measures for immigrants from Hubei Province may greatly reduce the risk of continued spread of the epidemic.

The complete chloroplast genome of the threatened Dipentodon sinicus (Dipentodontaceae)

Dipentodon is a monotypic genus of Dipentodontaceae and the only species, Dipentodon sinicus, is scattered in southwest China as well as adjacent Myanmar, northeast India and northern Vietnam. This species was evaluated as vulnerable in ‘China Species Red List’. Here, we assembled and characterized the complete chloroplast (cp) genome of D. sinicus using Illumina sequencing data for the first time. The complete cp genome was 158,795 bp in length, consisting of a pair of inverted repeats of 26,587 bp, a large single-copy region of 87,233 bp and a small single-copy region of 18,388 bp. The genome encoded 113 unique genes, including 79 protein-coding genes, 30 tRNA genes and four rRNA genes. Phylogenetic analysis based on 16 complete cp genome sequences indicated that D. sinicus is a member of Huerteales, consistent with its position in the latest classification of flowering plants (AGP IV).

Service demand and capacity of HIV prevention of mother-to-child transmission in Guangdong / 中华疾病控制杂志

Objective To analyze the service demand and capacity for preventing mother-to-child transmission(MTCT) of acquired immune deficiency syndrome in Guangdong, as well as to find the weakness in the work. Methods The relevant data of service demands and intervention capacity of human acquired immunodeficiency virus(HIV)-infected maternal and their babies from 2014 to 2017 was collected, and SPSS 21.0 software was employed to analyze the differences among the pearl river delta area, western area, mountainous area and eastern area, and to explore the correlation between regional midwifery institutions or personnel numbers and the rate of HIV MTCT. Results The education of HIV-infected maternal wasn’t high, generally, mainly in middle school (54.49%). The service demand of HIV-infected maternal was different in each area. The ethnic minorities outside the pearl river delta region (12.96%) and non-local living maternal in mountainous area (43.75%) were high. The proportion of maternal in the eastern area who didn’t know the route of HIV infection was also high (77.78%). The service capacity varies in each area, and the proportion (mountainous area: 41.67%, eastern area: 44.44%) of the confirmation time in intrapartum /postpartum was high among those from the mountains and eastern areas, which lead to poor intervention (the proportions of starting to antiretroviral treatment time later than 36 weeks or untreated in mountainous and eastern areas were 43.75% and 55.56%, respectively), as well as a high proportion of untreated maternal and exposed-infant (mountainous area: 2.08%, eastern area: 33.33%), and a higher percentage (11.11%) of breastfeeding in eastern area. There was no significant correlation between the number of midwifery institutions or healthcare personnel and the rate of HIV MTCT in different regions. Conclusions The mountainous and eastern areas are the weak of HIV MTCT in Guangdong Province, and later detection, less-standard intervention, and lower quality of information management are the weak steps in those areas. Targeted measures should be urgently developed to strengthen the training of the key areas to eliminate the regional differences of service capacity, which is essential to achieve the elimination of HIV MTCT in Guangdong.

Icariin promotes the proliferation of human periosteum cells and the underlying mechanism / 中国组织工程研究

BACKGROUND: Periosteal cells are precursors of osteoblasts and chondrocytes. Some studies have reported that bone morphogenetic protein-7 can be used to induce periosteal cell proliferation, but limited by the high cost. Phytoestrogen icariin-induced periosteal cell proliferation has provided a new direction for tissue engineering. OBJECTIVE: To investigate the effect of icariin on the proliferation of human periosteum cells and to analyze the underlying mechanism. METHODS:Human periosteal cells were cultured in vitro and seeded to the 24-well plate with the concentration of 103/well after third passage. Cell proliferation test: the cells were cultured in the cell culture medium (control group), or the culture medium containing different concentrations of icariin (10-1, 10-2and 10-3mg/L). Proliferation mechanism test: the cells were cultured in the cell culture medium (control group), or the culture medium containing different concentrations of icariin (10-1, 10-2and 10-3mg/L) plus estrogen receptor antagonist ICI182.780. The cell proliferation in each test was detected by MTT assay at 1, 2 and 3 days of culture. The effects of different concentrations of icariin on the levels of estrogen receptor α and β proteins in the periosteal cells were detected by western blot assay. RESULTS AND CONCLUSION:The proliferation of human periosteum cells in vitro was successful,and icariin with the concentrations of 10-1, 10-2and 10-3mg/L could significantly the cell proliferation (P < 0.05). However, this effect was blocked after ICI182780 addition (P < 0.05), and the levels of estrogen receptor α and β were upregulated. To conclude, icariin can enhance the proliferation of periosteal cells probably by upregualting the expression of estrogen receptor α and β.

Transcriptome profiling and analysis of Panax japonicus var. major / 中国中药杂志

The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.

Virological surveillance of pandemic (H1N1) 2009 virus and its genetic characteristics in Hunan Province, 2009-2011 / 病毒学报

To understand and master the dynamic variation of the pandemic influenza A (H1N1) 2009 in Hunan province from 2009 to 2011, and to know the genetic characteristics and drug resistance of the pandemic (H1N1) 2009 viruses. Throat swab specimens of influenza-like illness patients were collected from sentinel hospitals and tested for influenza by fluorescent PCR or virus isolation methods. Partial isolates were selected for sequencing. The sequences were used for phylogenetic analysis by MEGA 5. 05 software. From the 20th week of 2009 to the 52nd week of 2011, 17 773 specimens were tested. 3 831 specimens were influenza-positive with a positive rate of 21. 6%, of which 1 794 were positive specimens of pandemic (H1N1) 2009, accounting for 46. 8%00 of the influenza-positives. There were 2 epidemic peaks of pandemic (H1N1) 2009, which were in the 41st-53rd week of 2009 and the 1st-12nd week of 2011, respectively. The HA genes of 23 strains that were selected for sequencing had close relationship; the distribution of strains in the phylogenetic tree was basically in chronological order. The complete genome sequence analysis showed that all of 8 gene segments of 7 strains were homologous to the vaccine strain, and there was no gene reassortment. The HA amino acid sites of the 23 strains were highly similar to the vaccine strain (98. 2% - 100. 0% in homology), but all 23 strains had P83S, S203T and 1321V mutations. The 222 site mutation that may lead to enhanced virulence was found in the A/Hunan/YQ30/2009 strain. The mutation was D222E. There was no oseltamivir resistance mutation found in all strains. The pandemic (H1N1) 2009 in Hunan province from 2009 to 2011 had a bimodal distribution. There was no large-scale variation of virus genes. The clinical use of oseltamivir was still effective. Key words: Pandemic (H1N1) 2009; Surveillance; Genetic characteristics

The influence of familial factors on post rabies exposure vaccination in rural habitants / 中华流行病学杂志

Objective To understand the current situation of potential exposure to rabies among the rural habitants in Hunan province,and to study the impact related to familial factors on post rabies exposure vaccination.Methods In total,40 villages from 20 townships of 4 counties were selected by multistage sampling method.Study samples were selected from these villagers and familial basic information and vaccination post rabies exposures were recorded through questionnaires.Data were statistically analyzed by SPSS 17.0.Results Among 3042 villagers from 864 households being surveyed,124 person-time exposures were found from January,2009 to October,2010,with a total exposure rate as 4.08％,and the annual average exposure rate as 2.33％.Data from univariante analysis showed that the rates on post rabies exposure vaccination were statistically correlated with the following four factors:knowledge on the score of rabies prevention (x2 =8.260,P =0.042),whether being involved in the new type of rural cooperative medical care (P =0.035),family disposable cash income in the year of 2009 (x2 =10.831,P =0.031),distance between the households and the health facilities in towns and townships (x2 =9.071,P=0.033).Results from logistic regression analysis indicated that the score of knowledge on rabies prevention (OR=1.420,95％CI:1.055-1.905) and the annual disposable cash income of the family in 2009 (OR=1.480,95％ CI:1.044-2.098) were independent factors that influencing the rabies vaccination.Conclusion Strengthening the education programs on rabies prevention in rural habitants and increasing family income were feasible way to increase the rate of rabies vaccination in rural areas of Hunan province.

Analysis of genetic features of influenza B virus in Hunan province from 2007 to 2010 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the gene variations of influenza B virus isolated in Hunan province from 2007 to 2010.</p><p><b>METHODS</b>A total of 42 strains of influenza B virus,which were isolated in the Influenza Surveillance Network Laboratories in Hunan province between year 2007 and 2010, were selected for the study. The hemagglutinin 1 (HA1) and neuraminidase (NA) genes of the selected strains were amplified by RT-PCR, and the sequence of the purified product were detected and homologically compared with the sequence of influenza vaccine strains isolated from Northern Hemisphere by WHO during the same period. In addition, the phylogenetic trees were constructed to characterize the molecular features.</p><p><b>RESULTS</b>In the Victoria branch of the HA1 gene phylogenetic tree, the strains isolated from year 2007 to 2009 were included in the V1 sub-branch, as well as the vaccine strain Malaysia/2506/2004; the strains isolated in year 2010 were involved in the V2 sub-branch, similar to the vaccine strains Brisbane/60/2008. In the Yamagata branch,the strains isolated in year 2007 were in the Y1 sub-branch,different from the strains isolated between year 2008 and 2010, which were in the Y2 sub-branch, instead. All virus in NA gene phylogenetic tree were included in the Yamagata branch, indicated their Yamagata origin. The genetic sequence analysis of the 7 strains isolated in year 2010 revealed that the viruses were classified as genotype 2 and genotype 15. The results of homological comparison between HA1 molecule and the influenza vaccine strains recommended by WHO were as below: Victoria lineage, 98.6% - 99.1% in 2007, 98.6% - 99.1% in 2008, 98.1% - 99.1% in 2009, and 97.6% - 99.1% in 2010; and Yamagata lineage, 97.9% - 98.5% in 2007, 97.9% - 98.5% in 2009 and 97.9% - 98.2% in 2010. The major mutations of the strains isolated in year 2007 were found in sites R48K, K88R, P108A, D197N and S230G. While the major mutations of the strains isolated between year 2009 and 2010 were sited in K88R, S150I, N166Y, D197N and S230G.</p><p><b>CONCLUSION</b>The prevalent influenza B virus isolated in Hunan province from 2007 to 2010 has mutated and evolved continuously.</p>

Application of mesenchymal stem cells as a vehicle to deliver replication-competent adenovirus for treating malignant glioma / 癌症

Although gene therapy was regarded as a promising approach for glioma treatment, its therapeutic efficacy was often disappointing because of the lack of efficient drug delivery systems. Mesenchymal stem cells(MSCs) have been reported to have a tropism for brain tumors and thus could be used as delivery vehicles for glioma therapy. Therefore, in this study, we attempted to treat glioma by using MSCs as a vehicle for delivering replication-competent adenovirus. We firstly compared the infectivity of type 3, type 5, and type 35 fiber-modified adenoviruses in MSCs. We also determined suitable adenovirus titer in vitro and then used this titer to analyze the ability of MSCs to deliver replication-competent adenovirus into glioma in vivo. Our results indicated that type 35 fiber-modified adenovirus showed higher infectivity than did naked type 3 or type 5 fiber-modified adenovirus. MSCs carrying replication-competent adenovirus significantly inhibited tumor growth in vivo compared with other control groups. In conclusion, MSCs are an effective vehicle that can successfully transport replication-competent adenovirus into glioma, making it a potential therapeutic strategy for treating malignant glioma.

Serological survey of pandemic influenza A (H1N1) virus infection among population in Hunan Province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To understand the infection condition and analytical methods of Influenza A (H1N1) virus in the population of Hunan Province during different periods.</p><p><b>METHODS</b>Quick surveys on the positive rate of Influenza A (H1N1) virus hemagglutination inhibition (HI) test have been conducted for 5 times successively from November 2009 to March 2010 in 14 medical and health institutions of Changsha city, whose results were then compared with those from the sampling surveys of whole Hunan province.</p><p><b>RESULTS</b>2131 subjects were involved in this study; the total population standardized rates of antibody positive investigated for 5 times were 9.32% , 14.62%, 31.08%, 28.43% and 22.80% respectively; the population of 6-17-years-old has the highest rate of antibody positive; only 9.84% of the antibody positive subjects attributed to vaccine inoculation; there was no significant difference in the standardized positive rates between the quick serological surveys and the corresponding sampling survey of Hunan province (P > 0.05).</p><p><b>CONCLUSION</b>The positive rate of A (H1N1) virus antibody reached the peak in late January 2010; quick investigations in small region could be used to evaluate the infection prevalence during pandemic of infectious diseases.</p>