LDH-A-Modulation and the Variability of LDH Isoenzyme Profiles in Murine Gliomas: A Link with Metabolic and Growth Responses.

Three murine glioma cell lines (GL261, CT2A, and ALTS1C1) were modified to downregulate the expression of the murine LDH-A gene using shRNA, and compared to shRNA scrambled control (NC) cell lines. Differences in the expression of LDH-A and LDH-B mRNA, protein and enzymatic activity, as well as their LDH isoenzyme profiles, were observed in the six cell lines, and confirmed successful LDH-A KD. LDH-A KD (knock-down) resulted in metabolic changes in cells with a reduction in glycolysis (GlycoPER) and an increase in basal respiratory rate (mitoOCR). GL261 cells had a more limited ATP production capacity compared to CT2A and ALTS1C1 cells. An analysis of mRNA expression data indicated that: (i) GL261 LDH-A KD cells may have an improved ability to metabolize lactate into the TCA cycle; and (ii) that GL261 LDH-A KD cells can upregulate lipid metabolism/fatty acid oxidation pathways, whereas the other glioma cell lines do not have this capacity. These two observations suggest that GL261 LDH-A KD cells can develop/activate alternative metabolic pathways for enhanced survival in a nutrient-limited environment, and that specific nutrient limitations have a variable impact on tumor cell metabolism and proliferation. The phenotypic effects of LDH-A KD were compared to those in control (NC) cells and tumors. LDH-A KD prolonged the doubling time of GL261 cells in culture and prevented the formation of subcutaneous flank tumors in immune-competent C57BL/6 mice, whereas GL261 NC tumors had a prolonged growth delay in C57BL/6 mice. In nude mice, both LDH-A KD and NC GL261 tumors grew rapidly (more rapidly than GL261 NC tumors in C57BL/6 mice), demonstrating the impact of an intact immune system on GL261 tumor growth. No differences between NC and KD cell proliferation (in vitro) or tumor growth in C57BL/6 mice (doubling time) were observed for CT2A and ALTS1C1 cells and tumors, despite the small changes to their LDH isoenzyme profiles. These results suggest that GL261 glioma cells (but not CT2A and ALTS1C1 cells) are pre-programmed to have the capacity for activating different metabolic pathways with higher TCA cycle activity, and that this capacity is enhanced by LDH-A depletion. We observed that the combined impact of LDH-A depletion and the immune system had a significant impact on the growth of subcutaneous-located GL261 tumors.

Genetic and Drug Inhibition of LDH-A: Effects on Murine Gliomas.

The effects of the LDH-A depletion via shRNA knockdown on three murine glioma cell lines and corresponding intracranial (i.c.) tumors were studied and compared to pharmacologic (GNE-R-140) inhibition of the LDH enzyme complex, and to shRNA scrambled control (NC) cell lines. The effects of genetic-shRNA LDH-A knockdown and LDH drug-targeted inhibition (GNE-R-140) on tumor-cell metabolism, tumor growth, and animal survival were similar. LDH-A KD and GNE-R-140 unexpectedly increased the aggressiveness of GL261 intracranial gliomas, but not CT2A and ALTS1C1 i.c. gliomas. Furthermore, the bioenergetic profiles (ECAR and OCR) of GL261 NC and LDH-A KD cells under different nutrient limitations showed that (a) exogenous pyruvate is not a major carbon source for metabolism through the TCA cycle of native GL261 cells; and (b) the unique upregulation of LDH-B that occurs in GL261 LDH-A KD cells results in these cells being better able to: (i) metabolize lactate as a primary carbon source through the TCA cycle, (ii) be a net consumer of lactate, and (iii) showed a significant increase in the proliferation rate following the addition of 10 mM lactate to the glucose-free media (only seen in GL261 KD cells). Our study suggests that inhibition of LDH-A/glycolysis may not be a general strategy to inhibit the i.c. growth of all gliomas, since the level of LDH-A expression and its interplay with LDH-B can lead to complex metabolic interactions between tumor cells and their environment. Metabolic-inhibition treatment strategies need to be carefully assessed, since the inhibition of glycolysis (e.g., inhibition of LDH-A) may lead to the unexpected development and activation of alternative metabolic pathways (e.g., upregulation of lipid metabolism and fatty-acid oxidation pathways), resulting in enhanced tumor-cell survival in a nutrient-limited environment and leading to increased tumor aggressiveness.

[Epidemiological investigation of two leptospirosis death cases in Guizhou Province].

Objective: To conduct an epidemiological investigation of two leptospirosis death cases reported in Guizhou Province in 2014. Methods: The information of the patients were investigated and analyzed. The serological detection, samples of the two patients was detected using ELISA and microscopic agglutination test (MAT). Leptospira carrier status of murine host animal in the living environment of the two patients was investigated in October and November of 2014. Leptospires in the kidney were cultured and isolated, the isolates were identified using Leptospira specific PCR and further identified with serogroup specific PCR and the conventional MAT. The relativity between the carrier status of murine and the death cases of human leptospirosis was analyzed. Results: The two death cases of human leptospirosis came from Liping County and the clinical symptoms were consistent with the diagnosis criteria for Leptospirosis. The results of ELISA detection showed that the anti-Leptospira antibody was positive for one of the death cases, MAT identified the serum reacted with sera-group icterohaemorrhagiae Leptospira, while the serum sample of the other case was failed to perform antibody detection due to hemolysis. 1 600 traps were placed in the living environment of the two death cases and 183 murine rodents were trapped. The murine density was 11.44% (183/1 600); 40 leptospirea suspected strains were isolated and all of them were isolated from Apodemus agrarius. The positive rate was 21.86% (40/183); 95 Apodemus agrarius were trapped and the murine density was 5.93% (95/1 600). Species specific PCR identified all the 40 strains as Leptospire. Serogroup specific PCR further identification showed that they were iterohaemorrahgiae serogroup Leptospria. interrogans. Conclusion: Anti-iterohaemorrahgiae Leptospira antibody was detected from one of the two patients. 40 strains of iterohaemorrahgiae serogroup Leptospira interrogans were isolated and all of them were isolated from Apodemus agrarius in the living environment and the serogroup of the Leptospira matched with the serological detection results from patients, which indicated that the two death cases were caused by the infection of iterohaemorrahgiae serogroup Leptospira interrogans, and Apodemus agrarius were the potential source of infection.

Epidemiological investigation of two leptospirosis death cases in Guizhou Province / 中华预防医学杂志

Objective@#To conduct an epidemiological investigation of two leptospirosis death cases reported in Guizhou Province in 2014.@*Methods@#The information of the patients were investigated and analyzed. The serological detection, samples of the two patients was detected using ELISA and microscopic agglutination test (MAT). Leptospira carrier status of murine host animal in the living environment of the two patients was investigated in October and November of 2014. Leptospires in the kidney were cultured and isolated, the isolates were identified using Leptospira specific PCR and further identified with serogroup specific PCR and the conventional MAT. The relativity between the carrier status of murine and the death cases of human leptospirosis was analyzed.@*Results@#The two death cases of human leptospirosis came from Liping County and the clinical symptoms were consistent with the diagnosis criteria for Leptospirosis. The results of ELISA detection showed that the anti-Leptospira antibody was positive for one of the death cases, MAT identified the serum reacted with sera-group icterohaemorrhagiae Leptospira, while the serum sample of the other case was failed to perform antibody detection due to hemolysis. 1 600 traps were placed in the living environment of the two death cases and 183 murine rodents were trapped. The murine density was 11.44% (183/1 600); 40 leptospirea suspected strains were isolated and all of them were isolated from Apodemus agrarius. The positive rate was 21.86% (40/183); 95 Apodemus agrarius were trapped and the murine density was 5.93% (95/1 600). Species specific PCR identified all the 40 strains as Leptospire. Serogroup specific PCR further identification showed that they were iterohaemorrahgiae serogroup Leptospria. interrogans.@*Conclusion@#Anti-iterohaemorrahgiae Leptospira antibody was detected from one of the two patients. 40 strains of iterohaemorrahgiae serogroup Leptospira interrogans were isolated and all of them were isolated from Apodemus agrarius in the living environment and the serogroup of the Leptospira matched with the serological detection results from patients, which indicated that the two death cases were caused by the infection of iterohaemorrahgiae serogroup Leptospira interrogans, and Apodemus agrarius were the potential source of infection.

Evolution characteristics of hemorrhagic fever with renal syndrome in Huangdao and Jiaonan of Qingdao City, Shandong Province from 1979 to 2014 / 中华地方病学杂志

Objective To investigate the influence of the variations of an epidemic focus on hemorrhagic fever with renal syndrome (HFRS) so as to provide a basis for effective control of HFRS.Methods HFRS epidemic data from the rapid development of urbanization of Huangdao District and the general development of Jiaonan City were collected and analyzed retrospectively from 1979 to 2014.And the HFRS cases,incidences,host animals and Hantavirus surveillances were compared.Results The three epidemic peaks occurred from 1979 to 2014 in Huangdao District and Jiaonan City,they were in the mid-1980s (1983-1987),the late 1990s (1995-1999) and around 2012 (2010-2014),and 954,80,37 and 2 506,614,432 cases were reported,respectively,in Huangdao District and Jiaonan City.The three peak years were 1986,1999 and 2012.And the annual incidences of Huangdao District were 385.73/100 000,15.64/100 000 and 2.51/100 000,respectively.The annual incidences of Jiaonan City were 67.07/100 000,28.68/100 000 and 14.68/100 000,respectively.The three peaks obviously appeared double peaks [the autumn and winter (from Oct.to Dec.) and the spring (from Jan.to Feb.)] in Jiaonan City.And the first peaks in Huangdao District was in the autumn and winter (from Oct.to Dec.) and the spring (from Jan.to Feb.),the second and the third were only in the autumn and winter (from Oct.to Dec.).Eight kinds of host animals were detected in Jiaonan City from 2005 to 2014.They were house mouse [27.53％ (1 108/4 024)],brown rat [25.50％ (1 026/ 4 024)],striped field mouse [14.84％ (597/4 024)],black rat [10.74％ (432/4 024)],hamsters [11.01％ (443/4 024)],shrew [8.72％ (351/4 024)],back grain hamster [1.02％ (41/4 024)] and club rat [0.65％ (26/4 024)].Two kinds of host animals were detected in Huangdao District.They were house mouse [57.14％ (16/28)] and brown rat [42.86％ (12/28)].The capture rates in Jiaonan City were higher than those of Huangdao District.The capture rate in 2012 was 8.04％ (855/10 638) and the capture rates in Huangdao District were all lower than 0.84％.The total detection rate in Jiaonan City was 2.81％ (113/4 024).And there was no positive detection for ten years in Huangdao District.Conclusions HFRS epidemic intensities have receded gradually in Huangdao District and Jiaonan City.The epidemic in Huangdao District obviously presents a low epidemic condition after the first peak.The few kinds and low densities of host animals are resulting in the sustainable condition.The evolution of epidemic patterns of HFRS is related to the change of epidemic sources by the differences in urbanization between the two places.

Natural hosts infected with Babesia in Fujian Province, China / 中国人兽共患病学报

We explored the status of Babesia infection in natural hosts in Fujian Province.Rodents in Fujian Province were captured by the night trapping method during 2014-2016.Heart blood samples were collected.At the same time blood samples of ox,goats and dogs were also collected,from which the fragments of 18S rRNA gene of Babesia were amplified by polymerase chain reaction (PCR).Data on infection rate were analyzed with Chi-square or Fisher exact test to indicate statistical significance.Results showed that 5 917 cages were laid and 381 rodents were captured,density of rodent was 6.44％.The o verall Babesia infection rate in rodents was 7.61％,while infection rate in domesticated rodents was 1.68％,and the infection rate in wide rodents was 12.87％.The infection rate was higher in wild rodents than that in domesticated rodents,and the statistical analysis result revealed a significant difference.The infection rates in region of Central Fujian and Eastern Fujian was high,and no infection was found in Southern Fujian region.The statistical analysis result revealed a significant difference in infection rate among different regions.The infection rate of goats and dogs were determined to be 1.79 ％ and 0.55 ％,and no infection was found in ox.The infection rate was higher in rodents than that in other host animals,and the statistical analysis result revealed a significant difference.It is suggested that the rodents,especially the wide rodents are the main natural hosts of Babesia.

Natural hosts infected with Babesia in Fujian Province, China / 中国人兽共患病学报

We explored the status of Babesia infection in natural hosts in Fujian Province.Rodents in Fujian Province were captured by the night trapping method during 2014-2016.Heart blood samples were collected.At the same time blood samples of ox,goats and dogs were also collected,from which the fragments of 18S rRNA gene of Babesia were amplified by polymerase chain reaction (PCR).Data on infection rate were analyzed with Chi-square or Fisher exact test to indicate statistical significance.Results showed that 5 917 cages were laid and 381 rodents were captured,density of rodent was 6.44％.The o verall Babesia infection rate in rodents was 7.61％,while infection rate in domesticated rodents was 1.68％,and the infection rate in wide rodents was 12.87％.The infection rate was higher in wild rodents than that in domesticated rodents,and the statistical analysis result revealed a significant difference.The infection rates in region of Central Fujian and Eastern Fujian was high,and no infection was found in Southern Fujian region.The statistical analysis result revealed a significant difference in infection rate among different regions.The infection rate of goats and dogs were determined to be 1.79 ％ and 0.55 ％,and no infection was found in ox.The infection rate was higher in rodents than that in other host animals,and the statistical analysis result revealed a significant difference.It is suggested that the rodents,especially the wide rodents are the main natural hosts of Babesia.

Analysis of a monitoring results on animal plague in Meriones unguiculatus plague natural foci in Inner Mongolia from 2010 to 2014 / 中华地方病学杂志

Objective To analyze the monitoring results on plague in Meriones unguiculatus plague natural foci in Inner Mongolia from 2010 to 2014,to master the epidemic dynamics of the plague and to provide a basis for developing countermeasures.Methods The plague monitoring data in Meriones unguiculatus plague natural foci from 2010 to 2014 were collected;main host density,rate of dye fleas,flea body index and bacteriology were counted;serology detection was done and the epidemic situation was analyzed.Isolation and identification of Yersinia pestis were carried out through a 4-step test (microscopic examination,isolation and culture,phage lysis test and animal experiment).Serum samples were tested by indirect hemagglutination test.Results Within 5 years,21689 Meriones unguiculatus were captured overlap monitored areas of 7116 hm2 totally,the average rat density was 3.05/hm2;other small rodents were clipped 144352 times,3947 mice were captured,capture rate was 2.73％.Totally 26500 Meriones unguiculatus were checked,91 Meriones unguiculatus were infected with epidemic diseases,227 of 59 groups positive fleas were checked from cultured 51262 fleas of 13268 groups.Totally 5426 serum samples of Meriones unguiculatus were checked,5 copies were found positive,the positive rate was 0.09％.Conclusions Inner Mongolia Meriones unguiculatus plague is still active and spreading.We must enhance propaganda of the plague.Surveillance and emergency management should be strengthened to prevent a outbreak of the plague in human being.

Analysis of monitoring results on plague of rats in Lasiopodomys brandti plague natural foci in Inner Mongolia from 2001 to 2013 / 中华地方病学杂志

Objective To analysis the plague monitoring results on plague of rats in Lasiopodomys brandti plague natural foci of Xilin Gol Plateau in Inner Mongolia from 2001 to 2013,to master the dynamics of the plague epidemic,and to provide a basis for developing countermeasures.Methods Plague monitoring data from 2001 to 2013 in Lasiopodomys brandti plague natural foci were collected,main host density,rate of dye fleas,flea body index,bacteriology,serology and epidemic situation were studied.Results Within 13 years,10 153 Lasiopodomys brandti were captured overlapping a monitored area of 2 919.25 hm2,the average rat density was 3.48/hm2;other small rodents were captured 43 632 times,and 1 631 mice were captured,capture rate was 3.74％.Totally 22 752 host animals were checked by etiology,104 animals were infected with epidemic diseases,79 fleas of 31 groups positive fleas were checked from cultured 27 702 fleas of 6 437 groups.Totally 2 237 serum samples of Lasiopodomys brandti were checked using indirect hemagglutination (IHA),2 copies were found positive,the positive rate was 0.09％.Conclusion In Lasiopodomys brandti plague natural foci of Inner Mongolia,the plague bacteria infected host animals are still existed,future outbreak is possible;the monitoring and health education should be strengthened,in order to prevent the plague spreading to human being.

Blood meal analysis and virus detection in blood-fed mosquitoes collected during the 2006-2007 Rift Valley fever outbreak in Kenya.

BACKGROUND: Rift Valley fever (RVF) is a zoonosis of domestic ruminants in Africa. Blood-fed mosquitoes collected during the 2006-2007 RVF outbreak in Kenya were analyzed to determine the virus infection status and animal source of the blood meals. MATERIALS AND METHODS: Blood meals from individual mosquito abdomens were screened for viruses using Vero cells and RT-PCR. DNA was also extracted and the cytochrome c oxidase 1 (CO1) and cytochrome b (cytb) genes amplified by PCR. Purified amplicons were sequenced and queried in GenBank and Barcode of Life Database (BOLD) to identify the putative blood meal sources. RESULTS: The predominant species in Garissa were Aedes ochraceus, (n=561, 76%) and Ae. mcintoshi, (n=176, 24%), and Mansonia uniformis, (n=24, 72.7%) in Baringo. Ae. ochraceus fed on goats (37.6%), cattle (16.4%), donkeys (10.7%), sheep (5.9%), and humans (5.3%). Ae. mcintoshi fed on the same animals in almost equal proportions. RVFV was isolated from Ae. ochraceus that had fed on sheep (4), goats (3), human (1), cattle (1), and unidentified host (1), with infection and dissemination rates of 1.8% (10/561) and 50% (5/10), respectively, and 0.56% (1/176) and 100% (1/1) in Ae. mcintoshi. In Baringo, Ma. uniformis fed on sheep (38%), frogs (13%), duikers (8%), cattle (4%), goats (4%), and unidentified hosts (29%), with infection and dissemination rates of 25% (6/24) and 83.3% (5/6), respectively. Ndumu virus (NDUV) was also isolated from Ae. ochraceus with infection and dissemination rates of 2.3% (13/561) and 76.9% (10/13), and Ae. mcintoshi, 2.8% (5/176) and 80% (4/5), respectively. Ten of the infected Ae. ochraceus had fed on goats, sheep (1), and unidentified hosts (2), and Ae. mcintoshi on goats (3), camel (1), and donkey (1). CONCLUSION: This study has demonstrated that RVFV and NDUV were concurrently circulating during the outbreak, and sheep and goats were the main amplifiers of these viruses respectively.

Surveillance and control of Japanese encephalitis in Henan province, 1950-2008 / 中华流行病学杂志

oes of JE incidence in Henan province.