Objective To investigate the epidemiological characteristics of children infected by mycoplasma pneumoniae(MP)over the last five years in Kunming region.Methods Indirect immunofluorescence assay was used to detect blood MP-IgM of hospitalized patients from January 2003 to December 2007 in order to determine the age and gender distribution characteristics of MP infection and to investigate the epidemiological features of the five years' results.Results Infection incidences by MP in Kunming region over the last five years were 20.9%,14.3%,17.5%,15.7%and 19.5%,respectively.Statistical significance was found among the groups mentioned above(P＜0.01).The MP infection incidences in different age groups were 10.7%(～1 year old),20.5%(～3 years' old),21.5%(～6 years' old)and 21.7%(～14 years' old).MP infection incidences showed age and gender characteristics(P＜0.01).Infants showed lower NIP infection incidence and infection incidence was higher in female.The epidemiological investigation indicated that MP infection incidences varied from year to year(P＜0.01).The infection incidences of 2003 and 2007 by MP were higher than those of the other years.There were epidemiological differences in infection incidences by NIP over last five years(summer,autumn and winter).The MP infection incidences showed seasonal differences(P＜0.01).Conclusion Mycoplasma pneumoniae was the main local causative agent responsible for respiratory tract infection in children aged under one years' old in Kunming region.Over the last five years,there were two outbreaks in the local area,but with no seasonal regularity of epidemiology.

Objective To evaluate the efficacy of nucleoside analogues (NAs) antiviral therapy on clinical outcome for hepatitis B virus (HBV)-related primary hepatic carcinoma patients after hepatectomy. Methods The clinical data of 156 HBV-related primary hepatic carcinoma patients after hepatectomy were retrospectively analyzed..According to whether accepted postoperative antiviral treatment, all patients were divided into control group (n = 80)and observation group (n = 76). The serum HBV DNA capacity, recurrence-free survival (RFS)and overall survival (OS)were compared between two groups. Results One week, 1 month, 2 months and 3 months after operation , the serum HBV DNA capacity of observation group was significantly lower than that of control group(P < 0.05). One year, 3 years and 5 years after operation, intergroup comparison of RFS rate of both groups showed statistical significance (P < 0.05) and 1 year, 3 years and 5 years after operation, the difference of OS rate of both groups indicated statistical significance (P < 0.05). Conclusion Standard NAs antiviral treatment for HBV-related primary hepatic carcinoma patients after hepatectomy ,can improve prognosis and prolong survival time. The inhibition the HBV copy active may be its mechanism.

A bioprocess intensification strategy that combines both elicitation and in situ absorption was developed to improve the production of taxuyunnanine c (Tc) in cell suspension cultures of Taxus chinensis. When 100 micromol/L methyl jasmonate was added as an elicitor on Day 7, the Tc content and yield increased 3.6 and 3.3 times respectively, however the cell growth was reduced by 10%-30%. Significant improvement in Tc yield was observed when an absorbent XAD-7 was added on different time of the culture period. The optimum Tc yield was achieved when 100 g/L XAD-7 was added simultaneously with 100 micromol/L methyl jasmonate on Day 7. The maximum Tc yield of 477.4 mg/L was obtained on Day 21 of the culture, being 6.3-fold of the control and 1.9-fold of the 100 micromol/L methyl jasmonate treatment alone. In the combined treatment, 94% of the Tc produced was secreted outside of the cells and absorbed on XAD-7 absorbents. The results demonstrated that the process strategy combining elicitation and in situ absorption was effective to intensify the Tc biosynthesis via elicitation with the removal of product feedback inhibition via absorption, presenting a great potential in commercial applications.
Absorption ; Acetates ; pharmacology ; Acrylic Resins ; pharmacology ; Cell Culture Techniques ; methods ; Cyclopentanes ; pharmacology ; Oxylipins ; pharmacology ; Polystyrenes ; pharmacology ; Taxoids ; metabolism ; Taxus ; cytology

Objective To evaluate the BALB/c murine infective effects in different concentrations and different aerosol challenge times by Bordetella pertussis.Methods Four experiment groups according to different concentrations and different aerosol challenge times were designed.BALB/c murines were challenged by aerosol way.Group 1: 1010cfu/mL Bordetella pertussis challenge 15 min, group 2: 1010cfu/mL challenge 30 min, group 3: 109cfu/mL challenge 30 min, group 4: 1011cfu/mL challenge 30 min, using the normal saline challenge 30 min as control.At 0d,3d,7d,14d and 21d after challenge, the WBCs of all groups were measured and lung tissues were homogenized to calculate the bordetella pertussis clone in lung.Results After 3 days of challenge, WBCs in all groups were slightly increased.The WBCs of group 1, group 2, group 3 and group 4 were significantly increased after 7 days, with the average numbers of 8.52×109 per/L, 1.74×1010per/L, 1.15×1010per/L and 5×1010per/L, respectively.After 14 days, they were 1.77×1010per/L, 1.67×1010per/L, 1.27×1010per/L and 3.84×1010per/L respectively.WBCs in all groups were dramatically declined after 21 days.The WBC of negative control group had no obvious change during the whole process with the stable number of 3.4~7.0×109per/L.Bordetella pertussis were detected in lung of all experimental groups in each sampling point.The CFU in lung wase at peak at 7d or 14d after challenge, which was obviously decreased at 21d.Conclusion This aerosol challenge method can establish a bordetella pertussis infection mouse model successfully.

OBJECTIVETo construct a recombinant lentiviral vector that co-express green fluorescent protein (GFP) and FoxM1 shRNA and establish a prostate cancer cell line with stable FoxM1 down-regulation.

METHODSThree interfering sequences targeting FoxM1 were designed and inserted into the lentiviral vector pHBLV-U6-ZsGreen-Puro. After identification by DNA sequencing, the lentiviral vectors carrying Foxm1 shRNA were packaged in 293 cells. The lentiviral particles were collected to infect human prostate cancer DU-145 cells, and the transfection efficiency was observed under fluorescence microscope; the interference efficiency was assessed using real-time PCR. DU-145 cells with stable FoxM1 down-regulation were screened with puromycin, and the expression level of FoxM1 was detected by Western blotting and the cell growth was observed using MTT assay. The stably transfected cells were examined for cell apoptosis and cell clone formation capacity with flow cytometry and colony formation assay.

RESULTSDNA sequencing demonstrated successful construction of the 3 FoxM1 shRNA lentivirus vectors. Real-time PCR showed a high interference efficiency of FoxM1 shRNA1 vector, which resulted in obvious down-regulation of FoxM1 in DU-145 cells. Western blotting showed that the expression of FoxM1 protein was decreased in FoxM1 shRNA1 lentivirus-transfected cells, which displayed a suppressed cell proliferation, increased apoptosis rate, and attenuated clonogenic ability.

CONCLUSIONWe have successfully established a prostate cancer cell model with stable FoxM1 down-regulation, which shows lowered proliferative and clonogenic activities with increased cell apoptosis.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Forkhead Box Protein M1 ; Forkhead Transcription Factors ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Lentivirus ; Male ; Prostatic Neoplasms ; genetics ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Transfection

Taxus suspension cell culture has the potential to provide a sustainable source of anticancer drug paclitaxel (Taxol) and other taxoids. In the cell culture of Taxus chinensis, Taxuyunnanine C (Tc) is the primary taxoid. To design a rational strategy for redirecting the precursor fluxes from other taxoids into paclitaxel production, we employed Real-time Quantitative PCR (RQ-PCR) to understand the dynamic profiling of key biosynthetic pathway genes of palcitaxel and taxoids during the culture process. Six genes (TASY, TDAT, T5alphaH, TalphaH, T10betaH and T14betaH) were quantified under the process condition of double elicitation by 2,3-dihydroxylpropanyl jasmonate (DHPJA) (100 micromol/L on day 7 and day 12), and sucrose feeding (20 g/L) on day 7. This process treatment led to a high accumulation of Tc at (554.46 +/- 21.28) mg/L 8 days after the first elicitation. Then 9 days after the second elicitation, Tc production was as high as (997.72 +/- 1.51) mg/L. The early pathway genes TASY and TDAT were significantly up-regulated by 182-fold and 98-fold, respectively for the first DHPJA elicitation and by 208-fold and 131-fold, respectively for the second elicitation. The induction occurred after each elicitation lasted for about 24 h before their abundances decreased. Things are somewhat different in the case of the other four genes T5alphaH, TalphaH, T10betaH and T14betaH. For gene TalphaH, it was highly up-regulated by 3061-fold for the first DHPJA elicitation and by 1016-fold for the second elicitation. For the other three genes T5alphaH, T10betaH, T14betaH, they were up-regulated by 13-fold, 38-fold and 20-fold, respectively for the first DHPJA elicitation and by 7-fold, 16-fold and 6-fold, respectively for the second elicitation. The RQ-PCR results showed that there is tight correlation between gene expression and Tc accumulation. Gene expression was in accordance with Tc yield. Elicitation could improve expression of six genes. While along with culture course, high expression of the genes weakened. Elicitation for the second time would promote high expression of the genes again.
Cell Culture Techniques ; Culture Media ; Cyclopentanes ; pharmacology ; Gene Expression Regulation, Plant ; Oxylipins ; pharmacology ; Plant Growth Regulators ; pharmacology ; Sucrose ; pharmacology ; Taxoids ; metabolism ; Taxus ; genetics ; metabolism ; Transcriptome

Objective To construct a recombinant lentiviral vector that co-express green fluorescent protein (GFP) and FoxM1 shRNA and establish a prostate cancer cell line with stable FoxM1 down-regulation. Methods Three interfering sequences targeting FoxM1 were designed and inserted into the lentiviral vector pHBLV-U6-ZsGreen-Puro. After identification by DNA sequencing, the lentiviral vectors carrying Foxm1 shRNA were packaged in 293 cells. The lentiviral particles were collected to infect human prostate cancer DU-145 cells, and the transfection efficiency was observed under fluorescence microscope; the interference efficiency was assessed using real-time PCR. DU-145 cells with stable FoxM1 down-regulation were screened with puromycin, and the expression level of FoxM1 was detected by Western blotting and the cell growth was observed using MTT assay. The stably transfected cells were examined for cell apoptosis and cell clone formation capacity with flow cytometry and colony formation assay. Results DNA sequencing demonstrated successful construction of the 3 FoxM1 shRNA lentivirus vectors. Real-time PCR showed a high interference efficiency of FoxM1 shRNA1 vector, which resulted in obvious down-regulation of FoxM1 in DU-145 cells. Western blotting showed that the expression of FoxM1 protein was decreased in FoxM1 shRNA1 lentivirus-transfected cells, which displayed a suppressed cell proliferation, increased apoptosis rate, and attenuated clonogenic ability. Conclusion We have successfully established a prostate cancer cell model with stable FoxM1 down-regulation, which shows lowered proliferative and clonogenic activities with increased cell apoptosis.

Objective To construct a recombinant lentiviral vector that co-express green fluorescent protein (GFP) and FoxM1 shRNA and establish a prostate cancer cell line with stable FoxM1 down-regulation. Methods Three interfering sequences targeting FoxM1 were designed and inserted into the lentiviral vector pHBLV-U6-ZsGreen-Puro. After identification by DNA sequencing, the lentiviral vectors carrying Foxm1 shRNA were packaged in 293 cells. The lentiviral particles were collected to infect human prostate cancer DU-145 cells, and the transfection efficiency was observed under fluorescence microscope; the interference efficiency was assessed using real-time PCR. DU-145 cells with stable FoxM1 down-regulation were screened with puromycin, and the expression level of FoxM1 was detected by Western blotting and the cell growth was observed using MTT assay. The stably transfected cells were examined for cell apoptosis and cell clone formation capacity with flow cytometry and colony formation assay. Results DNA sequencing demonstrated successful construction of the 3 FoxM1 shRNA lentivirus vectors. Real-time PCR showed a high interference efficiency of FoxM1 shRNA1 vector, which resulted in obvious down-regulation of FoxM1 in DU-145 cells. Western blotting showed that the expression of FoxM1 protein was decreased in FoxM1 shRNA1 lentivirus-transfected cells, which displayed a suppressed cell proliferation, increased apoptosis rate, and attenuated clonogenic ability. Conclusion We have successfully established a prostate cancer cell model with stable FoxM1 down-regulation, which shows lowered proliferative and clonogenic activities with increased cell apoptosis.

Objective To investigate the effects of different concentrations and adsorption methods of aluminum hydroxide adjuvant produced by different manufacturers on the immunogenicity of the diphtheria-tetanus-acellular pertussis and inactivated poliovirus combined vaccine ( DTaP-sIPV) . Methods Five anti-gens of DTaP were adsorbed onto different concentrations (0. 42 mg/ml, 0. 47 mg/ml and 0. 52 mg/ml) of aluminum hydroxide from different manufacturers through sequential and separate adsorption. Adsorbability, anti-pertussis toxin ( PT)/filamentous hemagglutinin ( FHA)/pertactin ( PRN)/diphtheria toxoid ( DT)/tet-anus toxoid ( TT) antibodies and the potency of vaccines were detected. Results The adsorbability of alu-minum hydroxide adjuvant slightly decreased with the reduction of concentration. No significant difference in potency and antibody level was observed between sequential and separate adsorption. Moreover, no signifi-cant difference in antibody level was observed between vaccines prepared with aluminum hydroxide adjuvant produced by General Chemical Corp and our institute. Conclusion Aluminum hydroxide adjuvant produced by our institute at the concentration of 0. 52 mg/ml and separate adsorption method are suitable for prepara-tion of DTaP-sIPV.

Objective To analyze the literature related to ketogenic diet based on the Web of Science(WOS)database and clarify the research hotspots and trends in this field.Methods The literature related to ketogenic diet was retrieved from the core database of WOS from January 1,2013 to December 31,2022.Bibliometric methods and CiteSpace software were applied to statistically ana-lyze the literature characteristics,including countries,regions,research institutions,publishing jour-nals,and keywords.Results A total of 4,548 valid articles were retrieved in this study,including 3,317 original articles and 1,231 reviews.The number of publications in the field of ketogenic diet research showed an overall increasing trend from 2013 to 2022,reaching the peak in 2021(756 arti-cles).The annual citation frequency also exhibited a yearly increasing trend,reaching a peak of 25,643 citations in 2022.The country with the highest number of publications was the United States(1,695 articles,accounting for 37.27％),and the research institution with the most articles was Johns Hopkins University(88 articles,accounting for 1.93％).The majority of research institutions were universities.Nutrients journal had the highest number of published articles(241 articles,accounting for 5.30％),while Epilepsia journal had the highest average citation frequency per article(33.19 ci-tations).Among the top 10 most cited articles,five focused on the metabolism and mechanism of ac-tion of ketone bodies in disease treatment,and four were related to epilepsy research.The research hotspots included the application and efficacy evaluation of ketogenic diet in the treatment of epilepsy and obesity,common regulatory mechanisms such as fatty acid oxidation,metabolism,oxidative stress,and Warburg effect of ketone bodies,and the role of gut microbiota in mediating the effects of ketogenic diet.The research trend pointed towards the application prospects of ketogenic diet in heart failure.Conclusion Ketogenic diet has been applied in the treatment of various diseases,and its global attention continues to grow.Future research hotspots should focus on clinical trials to verify its clinical efficacy and tolerability.