Serial anti-tuberculous immune responses during the follow-up of patients with tuberculous pleurisy.

Little is known about the decay kinetics of interferon (IFN)-γ response and its influencing factors in tuberculous pleurisy. We enrolled thirty-two patients with tuberculous pleurisy prospectively and followed up at month 0, 6, and 9, at which time peripheral venous blood was drawn for interferon gamma release assay (IGRA) by means of QuantiFERON-TB Gold In-Tube (QFT-GIT). Demographic and clinical data were captured. To identify significant predictive factors influencing the IFN-γ response, multiple linear regression analyses were performed. Percentage of CD4+, CD8+, Vγ2Vδ2 T cells and Treg cells were measured by flow cytometry. The percentage of QFT-GIT-positive patients at baseline, month 6 and month 9 were 96.9% (30/32), 90.6% (29/32) and 84.4% (27/32), respectively. Quantitative IFN-γ response at baseline were significantly correlated with symptom duration (P = .003, R = 0.261) and age (P = .041, R = 0.132). Besides, the decreases of the IFN-γ response at month 6 and month 9 were positively correlated with the IFN-γ level at baseline. The dynamic tendency of the percentages of Treg cells was similar to the IFN-γ responses at each time-point. Quantitative IFN-γ response could be influenced by host immune status, instead of disease burden and anti-tuberculosis treatment. IGRA is probably not a useful biomarker of treatment efficacy in tuberculous pleurisy.

Curcumin-Protected PC12 Cells Against Glutamate-Induced Oxidative Toxicity.

Glutamate is a major excitatory neurotransmitter present in the central nervous system. The glutamate/cystine antiporter system x c- connects the antioxidant defense with neurotransmission and behaviour. Overactivation of ionotropic glutamate receptors induces neuronal death, a pathway called excitotoxicity. Glutamate-induced oxidative stress is a major contributor to neurodegenerative diseases including cerebral ischemia, Alzheimer's and Huntington's disease. Curcuma has a wide spectrum of biological activities regarding neuroprotection and neurocognition. By reducing the oxidative damage, curcumin attenuates a spinal cord ischemia-reperfusion injury, seizures and hippocampal neuronal loss. The rat pheochromocytoma (PC12) cell line exhibits many characteristics useful for the study of the neuroprotection and neurocognition. This investigation was carried out to determine whether the neuroprotective effects of curcumin can be observed via the glutamate-PC12 cell model. Results indicate that glutamate (20 mM) upregulated glutathione peroxidase 1, glutathione disulphide, Ca2+ influx, nitric oxide production, cytochrome c release, Bax/Bcl-2 ratio, caspase-3 activity, lactate dehydrogenase release, reactive oxygen species, H 2 O 2 , and malondialdehyde; and downregulated glutathione, glutathione reductase, superoxide dismutase and catalase, resulting in enhanced cell apoptosis. Curcumin alleviates all these adverse effects. Conclusively, curcumin can effectively protect PC12 cells against the glutamate-induced oxidative toxicity. Its mode of action involves two pathways: the glutathione-dependent nitric oxide-reactive oxygen species pathway and the mitochondria-dependent nitric oxide-reactive oxygen species pathway.

Hantaviruses in rodents and humans, Inner Mongolia Autonomous Region, China.

Surveys were carried out in 2003-2006 to better understand the epidemiology of hantaviruses in the Inner Mongolia Autonomous Region of China (Inner Mongolia). Hemorrhagic fever with renal syndrome (HFRS) was first reported in this region in 1955 and has been an important public health problem here since then. During 1955-2006, 8,309 persons with HFRS were reported in Inner Mongolia (average incidence rate 0.89/100,000), and 261 (3.14%) died. Before the 1990s, all HFRS cases occurred in northeastern Inner Mongolia. Subsequently, HFRS cases were registered in central (1995) and western (1999) Inner Mongolia. In this study, hantaviral antigens were identified in striped field mice (Apodemus agrarius) from northeastern Inner Mongolia and in Norway rats (Rattus norvegicus) from middle and western Inner Mongolia. Phylogenetic analysis of hantaviral genome sequences suggests that HFRS has been caused mainly by Hantaan virus in northeastern Inner Mongolia and by Seoul virus in central and western Inner Mongolia.

Effects of different glycerol feeding strategies on S-adenosyl-l-methionine biosynthesis by P(GAP)-driven Pichia pastoris overexpressing methionine adenosyltransferase.

Methionine adenosyltransferase (MAT) was overexpressed within Pichia pastoris employing the promoter of glyceraldehyde-3-phosphate dehydrogenase gene (P(GAP)), to biosynthesize S-adenosyl-l-methionine (SAM). Effects of five glycerol feeding tactics on MAT activity were first investigated. Strategies A-C were based on limited feeding correlated with dissolved oxygen (DO) at 50.0%, 25.0% and 0.0%, respectively. For strategies D and E, unlimited supplementation was executed by pulsed feeding mode. Gradual decline (2-0%) (w:v) of the residual glycerol level was shown between any two pulses in strategy D, while a nearly stable content (2%) throughout fed-batch cultivation with strategy E. With shifting strategies A-E in alphabetical order, gradual improvements of MAT activities were achieved, with the maximum of 9.05Ug(-1) dried biomass for strategy E, since the specific glycerol consumption rate (F(G)) ascended due to the elevated specific oxygen uptake rate (qO(2)). The success was ascribed to the enhancement of oxygen transfer rate (OTR), because 2% glycerol improved oxygen saturation content in broth (C*) and volumetric oxygen transfer coefficient (k(L)a). Strategy E also led to the highest values of ATP and biomass besides MAT. Consequently, the highest SAM yield and volumetric level were obtained at 0.058gg(-1) and 9.26gl(-1), respectively.

Isolation and genetic characterization of hantaviruses carried by Microtus voles in China.

To gain more insights into hantavirus distribution in China, Microtus fortis were caught in Jilin province and M. maximowiczii in the Inner Mongolia Autonomous Region. Hantavirus specific RNA was detected by RT-PCR in 3 out of 26 M. fortis and 5 out of 64 M. maximowiczii. Two hantaviruses (Fusong-Mf-682 and Yakeshi-Mm-59) were isolated successfully in cell culture and their S and M segment nucleotide sequences were determined. Phylogenetic analysis of the S and M segment sequences revealed that the Mf-originated strains from Fusong were closely related to Vladivostok hantavirus (VLAV) with 99% nucleotide identity, but differed from the Yakeshi-Mm strains, with an amino acid divergence of more than 8.8% for the N protein and 11.8% for the GnGc proteins. Yakeshi-Mm strains were closely related to the Khabarovsk hantavirus (KHAV) isolated earlier from M. fortis in Khabarovsk, with an amino acid sequence identity of more than 98.4% for the S segment and 95.6% for the M segment. On phylogenetic trees, Yakeshi-Mm strains clustered together with KHAV and Topografov virus (TOPV) carried by Lemmus sibiricus. The results suggest that the hantavirus carried by M. fortis in China belongs to VLAV type and should be considered as a distinct hantavirus species. They also suggest that M. fortis is the natural host of VLAV (including Fusong-Mf strains), whereas M. maximowiczii is the natural host of KHAV including Yakeshi-Mm strains. Thus, in addition to Hantaan, Seoul, Dabieshan and Puumala-like Hokkaido viruses, at least two other hantaviruses, namely KHAV and VLAV, are circulating in China.

[Epidemiological characteristics and the strategy of vaccination on hemorrhagic fever with renal syndrome in the last 10 years, in China].

OBJECTIVE: To study the epidemiological characteristics and vaccination strategy of hemorrhagic fever with renal syndrome (HFRS) in China, from 1998 to 2007. METHODS: National data on the prevalence of HFRS during 1998 to 2007 were collected and analyzed. Spatial, time and population distributions of HFRS cases were described. RESULTS: HFRS appeared to remain endemic in the country, while the majority of cases (accounting for 81.76% of total cases), were located in Shandong, Heilongjiang, Liaoning, Hebei, Shaanxi, Hunan, Jilin and Zhejiang provinces. The occurrence of HFRS appeared to be seasonal, with two peaks in spring season and autumn/winter season, with 10.00% of the cases occurred in these peak months. The incidence rates were at a relatively low level because of the continuous decline of HFRS in the recent years. With age distribution, 0.97% were younger than 10, 96.71% were between 10 and 70, 87.32% were between 15 and 60, and 2.31% were older than 70. The proportion of male patients accounted for three quarters. Around 70.00% of the cases were farmers. Young and middle-aged male farmers were at high risk. CONCLUSION: The descriptive study on epidemiological characteristics of HFRS in the recent years had provided scientific evidence for vaccination strategy, suggesting the key regions, sub-populations as well as types of vaccines be determined.

Detection of phylogenetically distinct Puumala-like viruses from red-grey vole Clethrionomys rufocanus in China.

In order to investigate whether Puumala virus (PUUV) or PUUV-like virus is present in China, Clethrionomys rufocanus and C. rutilus were captured in the Jilin province during the spring and autumn of 2002-2003 for detection of PUUV viral RNA by RT-PCR and confirmation of PUUV-positive antigens by an immunofluorescence assay. PUUV-positive RNA was identified in six out of 121 C. rufocanus but not in any of the 41 C. rutilus. Complete S and partial M sequences (nt 1,316-1,598 and 2,687-3,089) were amplified by RT-PCR directly from some of the antigen positive lung tissues and subjected to nucleic acid sequencing. It was found that the Chinese PUUV-like viruses were related most closely with the PUUV strains with 77.7-81.7% identity at the nucleotide level and 91.7-97% identity at the amino acid level for S segment, and with 77-78.8% identity at the nucleotide level and 91.5-92.6% identity at the amino acid level for the partial M segment (nt 1,316-1,598). Genetic analysis indicated that the Chinese PUUV-like viruses shared the highest level of identity with the viruses which circulate in C. rufocanus in the Far East region of Russia with 85.1-87.4% identity at the nucleotide level and 95.9% identity at the amino acid level for the partial M segment (nt 2,687-3,089), respectively. Phylogenetic analysis revealed that the Chinese PUUV-like viruses are distinct from those identified from Japan, South Korea, Europe or Russia. These results indicate that PUUV-like virus is present in China in addition to Hantaan, Seoul and Dabieshan viruses.

[An epidemiologle investigation of hantaviruses carried by rodent hosts in Hunan province].

OBJECTIVE: To explore the hantavirus infection and their genotype in rodents in Hunan. METHODS: Hantavirus antigens in the rat lungs from Hunan province were detected by immunofluorescence assay. Partial S and M segment in antigen-positive samples were amplified by RT-PCR, and then sequenced. The phyologenetic trees were constructed for the analysis of genetic characters of hantavirus. RESULTS: A total of 344 rats were trapped in the main epidemic area of Hunan province, and hantavirus antigens were found in 6 of the 344 rats( 1.74% ).The phylogenetic trees constructed by partial S segment( nt 620-990) or partial G2 segment (nt 2001- 2301) showed that the hantaviruses carried by Rattus norvegicus, R . flabipectus and R. rattoides from Xiangxiang district were genetic subtype SEOV4. The virus carried by R. norvegicus in Ningyuan district was phylogenetically different from the known SEOV. The hantavirus carried by Mus musculus from Shimen district was genetic subtype HTNV4. CONCLUSION: The hantaviruses in the main epidemic areas in Hunan province mainly belonged to SEOV, and R. flabipectus and R. rattoides carried the same genotype of SEOV as R. norvegicus.

Isolation and characterization of hantavirus carried by Apodemus peninsulae in Jilin, China.

To provide a better understanding of hantavirus epidemiology in China, Korean field mice (Apodemus peninsulae) and striped field mice (Apodemus agrarius) were captured in Jilin province, China, where haemorrhagic fever with renal syndrome (HFRS) is endemic. Hantavirus antigens were detected in eight of the 130 A. peninsulae individuals and in four of the 193 A. agrarius individuals by using an immunofluorescence assay. Partial S and M segments were amplified from all of the antigen-positive samples. Furthermore, two hantaviruses (CJAp89 and CJAp93) were isolated successfully in cell culture and the entire S and M segments were amplified from one of them (CJAp93). Phylogenetic analysis of these sequences (partial or complete) showed that hantaviruses carried by A. peninsulae and A. agrarius form two distinct lineages, although viruses carried by A. peninsulae are similar to those isolated previously from A. agrarius in China and from HFRS patients in Russia. However, the viruses detected in A. peninsulae in China are genetically different from those detected in A. peninsulae in other countries. These data suggest that A. peninsulae is also a natural host for HTNV in north-eastern China.

[Study on the epidemiological characteristics of hemorrhagic fever with renal syndrome in Inner Mongolia].

OBJECTIVE: In order to better understand the epidemiological features of Hantviruses in Inner Mongolia. METHODS: Epidemiological surveillance data during the period of the past 52 years were analyzed. An epidemiological survey was carried out in the main epidemic areas in 2005. RESULTS: A total of 8310 hemorrhagic fever with renal syndrome (HFRS) cases were reported in Inner Mongolia from 1955 to 2006, and distributed in 61 counties. HFRS cases were mainly distributed in the east part of Inner Mongolia before 1990. However, HFRS cases had occurred in the middle and western parts since 1990. Hulunbeier prefecture, from the eastern part of Inner Mongolia, had been the most severe area being hit by HERS since the first outbreak in 1955, with 7369 cases reported over the past 52 years, and accounted for 88.68% of the total cases in the whole autonomous region. Although no HFRS cases had been reported before 1999 in Bayannaoer which located in the western part of Inner Mongolia, a total of 95 cases were reported in 2005. Hantavirus antigens had been detected in 11 species of rodents so far,including Apodemus agrarius, Rattus norvegicus, Mus Musculus, Cricetulus barabensis, meriones meridianus, Microtus maximowiczii , Clethrionomys rutilus, Apodemus peninsulae, Phodopus roborvskii, Dipus sagitta and Allactaga sibirica. CONCLUSION: Results suggested that the epidemics might remain at a relatively high level in the years to come in Inner Mongolia. Furthermore, there might be other types of Hantaviruses in addition to the already identified Seoul viral type in this area.

[Isolation and characterization of Hantavirus carried by rodents in Huludao, Liaoning province].

OBJECTIVE: To investigate the Hantavirus infection and their genotype in rodents in Huludao. METHODS: Rodents were collected from the main epidemic areas to detect antigen of Hantavirus in rat lungs by indirect immunofluorescence assay. Antigen-positive samples were inoculated onto cultures of confluent Vero E6 cells for the isolation of virus. The genotypes of viruses in all antigen-positive samples were identified by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: 200 rats were collected in the main epidemic areas, and 11 Hantavirus-positive samples were tested. The positive rate of Hantavirus in rats was 5.5%. Three strains of Hantavirus were isolated in Vero E6 cell culture. Data from the phylogenetic trees constructed by partial S segment (620-999 nt) or partial G1 segment (180-580 nt) showed that the three isolates carried by rats from Huludao were all genetic subtype SEOV 3. Furthermore, the phylogenetic tree constructed by partial G2 segment (2003-2302 nt) divided SEOV strains into 7 genetic subtypes, and the three isolates were having a closer evolutionary relationship with isolates CP211, ch302 and dc501 from Beijing, and the isolates SD10 and SD227 form Shandong. CONCLUSION: Data indicated that the rate of carrying virus was high and the main genetic subtype of Hantavirus was S3 of Seoul virus in Huludao area.

[Genetics subtypes and distribution of Seoul virus in Henan].

OBJECTIVE: To investigate the sub-genotypes and distribution ot Seoul virus in Henan. METHODS: Rodents were collected in the major epidemic areas and rats lungs were studied by indirect immunofluorescence assay. Partial M and S segments were amplified with nested reverse transcription-polymerase chain reaction using Hantavirus genotype-specific primers, sequenced, analyzed and compared with other known sequences. RESULTS: The Hantavirus carried by Rattus norvegicus, Rattus flavipectus and Mus musculus were all belonged to Seoul virus in the main epidemic areas of Henan. We constructed two phylogenetic tree based on the partial M and S segment sequences while phylogenetic analysis distinguished three genetic subtypes (S1, S2 and S3). S1 and S3 were found main subtypes in Henan. CONCLUSION: The results indicated that the genetic subtypes of Hantavirus were complicated and widely distributed in Henan.

[NaCl stress increases the heat tolerance of PSII in leaves of rumex K-1 seedlings].

By measuring chlorophyll fluorescence, the effects of NaCl treatment on the maximal efficiency and heat tolerance of PSII were examined in leaves of Rumex seedlings. NaCl 200 mmol/L treatment had no effect on the maximal efficiency of PSII, but increased the heat tolerance of PSII in Rumex leaves. Compared with control leaves, the heat stress-induced decrease in F(v)/F(m) and the increase in F(k)/F(j) ratio were less in NaCl-treated leaves. In addition, the heat stress-induced decrease in photochemical quenching (q(P)), the efficiency of excitation energy capture by open PSII reaction center (F(v)' /F(m)') and the quantum yield of electron transport (PhiPSII) were less in NaCl-treated leaves. Moreover, the increase in the Q(B)-non-reducing PSII reaction center content was less in NaCl-treated leaves than in control leaves. Discussion was made on the possible mechanisms of the increase in the heat tolerance of PSII in NaCl-treated leaves.

Characterization of PSII photochemistry and thermostability in salt-treated Rumex leaves.

A study was conducted, using chlorophyll fluorescence, rapid fluorescence induction kinetics, and polyphasic fluorescence transients, to determine the effect of salt treatment and heat stress on PSII photochemistry in Rumex leaves. Salt treatment was accomplished by adding NaCl solutions of different concentrations ranging from 50 to 200 mmol/L. Heat stress was induced by exposing the plant leaves to temperatures ranging from 29 to 47 degrees C. The control plants were grown without NaCl treatment. The data acquired in this study showed that NaCl treatment alone had no effect on the maximal photochemistry of PSH or the polyphasic rise of chlorophyll fluorescence. However, the NaCl treatment modified heat stress on PSII photochemistry in Rumex leaves, which was manifested by a lesser heat-induced decrease in photochemical quenching (qP), efficiency of excitation energy capture by open PSII reaction centers (Fv'/Fm'), and quantum yield of PSII electron transport (phiPSII). The data also showed that NaCl treatment compromised the impact of heat stress on the capacity of transferring electrons from Q(A)- to Q(B). Furthermore, the NaCl treatment promoted heat resistance of O2-evolving complex (OEC). In summary, NaCl treatment enhanced the thermostability of PSII.

[Using geographic information system to study on the association between epidemic areas and main animal hosts of hemorrhagic fever with renal syndrome in China].

OBJECTIVE: To analyze the association between types of epidemic areas and proportion of main animal hosts of hemorrhagic fever with renal syndrome (HFRS) in China. METHODS: Data on serotype of HFRS patients from 122 regions and species as well as number of animal hosts from 41 national surveillance sites were collected and managed. The map of the types of epidemic areas from 1991-1999 in China was produced using inverse distance weighted technique in the ArcGIS 8.1 software. The map of the proportion of host animals was then overlapped on it and the surveillance sites of HFRS were clustered in terms of the proportion of host animals. RESULTS: Variance in spatial distribution of the types of epidemic areas was observed. Two epidemic area types, namely Hantaan-dominant mixed epidemic areas and Soul-dominant mixed epidemic areas seemed to have transferred from the north-east to south-west of China in turns. There were more species of HFRS animal hosts in fields than those of in residential areas. The diversity of rodent species in fields was higher than that of the residential areas which predominant harbored rodent species of 2-3 kinds. Types of surveillance sites classified by cluster analysis of proportion of main animal hosts were consistent with the serotypes of HFRS patients. CONCLUSION: Through employment of spatial analysis technique based on GIS databases, the HFRS types of main epidemic area in China could be well predicted by surveillance data.

[Typing of hemorrhagic fever with renal syndrome by cluster analysis].

OBJECTIVE: To make an inquiry into method of typing of hemorrhagic fever with renal syndrome (HFRS). METHOD: Average monthly rates were calculated on the basis of data from 1995 to 1999, then cluster analysis was carried out to type out endemic areas. RESULTS: Compared with the results of 36 surveillance spots from 1980 to 1992, twenty-four surveillance spots had the same results (66.7%). Twenty-three surveillance spots had the same results with the original data in 1999 (82.1%). CONCLUSION: HFRS incidences increased in spring or in summer, but decreased in autumn or in winter. Cluster analysis seemed to be a supplementary method in distinguishing the epidemic types for HFRS.

[Study on the application of geographic information system in spatial distribution of hemorrhage fever with renal syndrome in China].

OBJECTIVE: To analyze the spatial distribution of hemorrhagic fever with renal syndrome (HFRS) in China by geographic information system, and to draw up a map on HRFS risk areas. METHODS: A set of database was set up using the information collected and linked to electronic maps of China in a software ArcGIS 8.01 from 41 HFRS surveillance sites during 1995 - 1998. A HFRS spatial distribution model was developed using inverse distance weighted interpolation of ArcGIS's spatial analysis method. The normalized difference vegetation index (NDVI) in each HFRS surveillance site was extracted from SPOT4 satellite vegetation imagery. Correlation analysis was performed through SPSS 10.0 to analyze the association between NDVI and HFRS incidence, HFRS risk areas were mapped under different colors. RESULTS: Spatial distribution model from HFRS surveillance sites showed that HFRS foci mainly presented in the Heilongjiang River drainage, the middle and lower reaches of the Yellow River, the middle and lower reaches of the Yangtze River, and the Jinghang grant Canal-Huaihe River drainage. It was consistent with HFRS distribution map derived from national infectious disease reporting system. Correlation analysis indicated that HFRS incidence rates were significantly associated with NDVI (r = 0.417, P < 0.01). The HFRS risk areas was mapped according to NDVI of each surveillance site. CONCLUSION: It is promising to apply GIS technology in predication of the distribution of HFRS by establishing this prediction model.