Bis(ethyl-enediamine-κN,N')(nitrato-κO,O')cobalt(III) hydroxide nitrate.

The Co ion in the title salt, [Co(NO(3))(H(2)NCH(2)CH(2)NH(2))(2)](OH)(NO(3)), has oxidation state + III and is coordinated by four N atoms from two ethyl-enediamine mol-ecules and two O atoms from a nitrate anion in a distorted octa-hedral geometry. The charge of the complex cation is balanced by a hydroxide anion and a nitrate anion. The cations and anions are connected by N-H⋯O and O-H⋯O hydrogen bonds, resulting in a three-dimensional supra-molecular framework. There are two independent ion pairs with similar configurations in the unit cell. Both uncoordinated nitrate counter-anions are disordered.

Application of stereotactic biopsy for diagnosing intracranial lesions in patients with AIDS in China: Report of 7 cases.

RATIONALE: The aim of the study was to evaluate stereotactic biopsy for diagnosing intracranial lesions in patients with AIDS. PATIENT CONCERNS: Seven AIDS patients with an intracranial lesion who underwent stereotactic biopsy were included in this retrospective study (4 males and 3 females, 15 to 49 years old). The patients' disease history ranged from 1 month to 1 year. The samples were examined by hematoxylin-eosin (HE) staining and immunohistochemical examination. DIAGNOSES, INTERVENTIONS AND OUTCOMES: All patients were successfully sampled, and the histological results showed inflammation in 4 cases, toxoplasma gondii infection in 1 case, astrocytoma in 1 case, and abscess in 1 case. The clinical diagnosis included toxoplasma encephalitis (TE) in 2 cases, cryptococcus encephalitis in 2 cases, cytomegalovirus (CMV) encephalitis in 2 case, tubercular abscess in 1 case, astrocytoma in 1 case, and co-infection of TE with Cryptococcus infection in 1 patient. The clinical diagnosis was made according to the plasma and cerebrospinal fluid (CSF) laboratory testing, the imaging data and the histological findings. The diagnostic yield was 100%, and the post-operation morbidity was 14.3% (1/7) with an asymptomatic haemorrhage and seizure in 1 case. There was no operation-related mortality. Patients were followed up for 6 months to 6 years; 1 case fully recovered, 4 cases significantly improved in symptoms, and 2 died. LESSONS: Stereotactic biopsy is a safe and effective way of diagnosing intracranial lesions in patient with AIDS. It is helpful for the differential diagnosis and for choosing a suitable therapy. Due to the broad spectrum of nervous system abnormalities in AIDS, histological findings are very valuable. However, histology is not a unique tool for making a definite diagnosis, whereas the combination of molecular pathology and stereotactic biopsy should play a more important role in the future.

Construction and functional activity of a recombinant vector expressing rat glutamic acid decarboxylase 65.

OBJECTIVE: Glutamic acid decarboxylase 2 (GAD65) is a gamma-aminobutyric acid (GABA) synthetase. This study aimed to construct a recombinant lentivirus-rGAD65 (rLV-rGAD65) vector containing the cDNA of rat GAD65 (rGAD65) and assess its functional activity in vitro and in vivo. METHODS: cDNA of rGAD65 was amplified by RT-PCR and subcloned into the LV vector, forming the rLV-GFP-rGAD65 plasmid. The recombinant lentivirus particles (rLV-rGAD65) were packaged by the LV Helper-Free System and the titer was measured. Primary rat lung fibroblasts were transfected with rLV-rGAD65. The expression of rGAD65 in fibroblasts was detected by immunocytochemistry and western blot and the level of GABA in the medium was assessed by high-performance liquid chromatograph (HPLC). In vivo, rLV-rGAD65 was injected into the subthalamic nucleus (STN) of Sprague-Dawley rats using stereotaxic methods, and rGAD65 protein levels in the STN were assessed by immunohistochemistry and Western blot, while the GABA concentration in the substantia nigra pars reticulata (SNr) was assayed by HPLC. RESULTS: The sequence of rGAD65 cDNA was in accord with that in GenBank. The amino-acid sequence of rGAD65 had no mutations and the titer of rLV-rGAD65 reached 6.8 × 108/mL. The efficiency of infection of fibroblasts was 80%, and the concentration of GABA in the medium was (48.14 +/- 9.35) nmol/L. In vivo, rGAD65 expression was detected in the STN, and the concentration of GABA in the SNr increased from (5.95 +/- 1.09) to (12.44 +/- 3.79) nmol/g tissue. CONCLUSION: The recombinant LV-GFP-rGAD65 vector was successfully constructed. rLV-rGAD65-infected primary fibroblasts in vitro and the expressed rGAD65 catalyzed the formation of GABA from glutamic acid. In vivo, the concentration of GABA in the SNr was increased after rLV-rGAD65 injection into the STN.

[Molecular characterization of full-length genome of Japanese encephalitis virus isolated in Liaoning Province in 2008].

OBJECTIVE: To sequence and analyze the whole genome of Japanese encephalitis virus (JEV) isolated from mosquitoes in Liaoning province in 2008. METHODS: Using RT-PCR to amplify fragments with genome sequencing primer. The full-length genome was obtained by sequencing and splicing. The differentiation analysis for nucleotides, deduced amino acid sequence and phylogenetic tree was performed by the software of Clustal X (1.83), ATGC (V4), DNAStar, GENEDOC (3.2) and Mega (4.0). RESULTS: The whole genome of strain LN0828 possesses 10 965 nucleotides. An open reading frame from 97 to 10 392 including 10 296 nucleotides is capable of coding for a 3432 amino acid polyprotein. Comparison of strain LN0828 genomic sequence with those of 32 JEV isolates in GenBank showed that nucleotide sequence divergence ranges from 1.6% to 16.4%, which resulted in amino acid sequence divergence from 0.3% to 5.1%. In comparison with live attenuated vaccine stain SA14-14-2 in open reading frame, strain LN0828 has a total of 1186 nucleotide substitutions, 86 amino acid divergences. Based on phylogenetic analysis, the strain LN0828 belongs to the genotype I JEV. CONCLUSION: The whole genome of strain LN0828 is close to those of isolates from Liaoning in 2002 and 2007, which were grouped into genotype I JEV.

[Isolation and identification of arboviruses from mosquito pools in some regions of Liaoning province, China].

OBJECTIVE: To isolate and identify arboviruses from mosquito pools in some regions of Liaoning province. METHODS: Mosquitoes were collected from Shenyang, Yingkou, Panjin, Jinzhou and Dandong cities of Liaoning province in 2006. Viruses were isolated by inoculating the specimens onto C6/ 36 and BHK-21cells. The new isolates were identified using serological and molecular biological methods. RESULTS: 5410 mosquitoes were collected from the five cities in total. Three isolates produced CPE in C6/ 36 cell and five isolates produced CPE in both C6/36 and BHK-21 cell. Three isolates (LN0684, LN0688 and LN0689) were identified as Banna virus and one isolate (LN0636) was identified as Getah virus. Phylogenetic analysis showed that the three Banna virus strains were clustered into the same evolution branch as the other Chinese isolates. The identity of nucleotide sequence was between 91.2% and 94.7%, compared with other Banna virus strains. The new isolated Getah virus was clustered into the same branch with the strain of South Korea (swine). The identity of nucleotide sequence was 99.2%, when comparing with the strain of South Korea and was 95% to 99% with the strains from Russia, mainland of China and Taiwan region. Conclusion Eight virus isolates, including three Banna virus, one Getah virus and four unknown virus strains were isolated from mosquitoes in Liaoning province. Banna virus and Getah virus were reported for the first time in Liaoning province, while Getah virus showed the highest nucleotide homology with the South Korea strains.

[Genotype 1 JEV was isolated again from Liaoning Province, China, 2007].

OBJECTIVE: To isolate Japanese encephalitis virus (JEV) from mosquitoes collected in Liaoning province and analysis the genotype of new isolated JEV strains and the characters of nucleotide and amino acid in the E gene. METHODS: Collected mosquitoes in Dandong Liaoning Province in August, 2006. Virus isolation was using issue culture cells. Isolated viruses were identified by using serological and molecular methods. RESULTS: Two new JEV strains, LNDG07-02 and LNDG07-16, were isolated from 1500 mosquitoes were belonging to genotype 1. The identity of nucleotide and amino acid of E gene between new JEV strains and live attenuated vaccine strain SA14-14-2 were 87.8-88% and 97.2%, respectively. Total 11 amino acid sites were differences in E gene between new isolates and SA14-14-2. However, there were no differentiation between the new JEV strains and the isolates in Donggang 2002. CONCLUSION: Genotype 1 JEV was isolated again from Donggang, since the first isolation of this genotype in 2002. Genotype 1 JEV continues in existence in Donggang Liaoning Province.

Non-Redundant Roles for Interleukin-1α and Interleukin-1ß in Regulating Human IgG2.

BACKGROUND: Serum concentrations of immunoglobulin G2 (IgG2) are elevated in localized aggressive periodontitis (LAgP) patients, and secretory products of monocytes from LAgP patients enhance IgG2 responses of lymphocytes from healthy subjects. Furthermore, genes regulating production of interleukin (IL)-1 influence the risk for both aggressive periodontitis (AgP) and chronic periodontitis. These observations, and the fact that IgG2 dominates responses to carbohydrates from Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis, prompted the hypothesis that IL-1α, IL-1ß, and IL-RA may help regulate human IgG2 responses. METHODS: Human peripheral blood leukocytes (PBL) were stimulated in culture with pokeweed mitogen (PWM); the levels of available IL-1 gene products were manipulated; and the effect on IgG2 production was monitored. Manipulations of IL-1 were accomplished by adding specific neutralizing monoclonal antibodies or recombinant IL-1RA, IL-1α, or IL-1ß. RESULTS: Blocking the IL-1 receptor with IL-1RA or neutralizing IL-1α or IL-1ß with specific antibody dramatically suppressed IgG2 production (50% to 70%). Additional IL-1α did not compensate for neutralized IL-1ß, and additional IL-1ß did not compensate for neutralized IL-1α, suggesting the 2 monokines have separate roles in promoting IgG2. Furthermore, combinations of anti-IL-1α and anti-IL-1ß were more inhibitory than either antibody alone, and IL-1α and IL-1ß in combination appeared to work additively in promoting IgG2. Moreover, PBL cultures from a group of LAgP patients with high IgG2 levels had elevated levels of IL-1ß. CONCLUSION: IL-1α and IL-1ß appear to have critical and nonredundant roles in the generation and regulation of potent IgG2 responses, which appear to be important in human responses to carbohydrate-bearing bacteria. J Periodontol 2001;72:1332-1339.

Antibody of the IgG2 Subclass, Actinobacillus actinomycetemcomitans, and Early-Onset Periodontitis.

Susceptibility to early-onset periodontitis (EOP) appears to be attributable to a gene inherited in an autosomal dominant pattern. This explains why EOP clusters in families and why about half of the family members develop periodontal disease early in life. Manifestation of EOP is variable, with some patients having a localized form restricted to first molars and incisors (LJP) and others with a severe generalized form of periodontitis (SP). The extent and severity of disease is less in patients who are seropositive for Actinobacillus actinomycetemcomitans than in seronegative patients, and this relationship prompted the hypothesis that anti-A. actinomycetemcomitans helps limit disease. The dominant antibody is an IgG2 reactive with the serotypespecific carbohydrate. The incidence of the LJP form of EOP is about 10 times higher in blacks than in whites. Interestingly, blacks have higher levels of serum IgG2, a higher frequency of anti-A. actinomycetemcomitans antibody, and higher serum titers of IgG2 anti-A. actinomycetemcomitans which may help explain why the disease is localized. Studies in progress suggest that smoking reduces serum IgG2 levels in SP patients and is associated with more severe periodontal destruction. In marked contrast, IgG2 does not appear to be reduced in LJP patients who smoke, and smoking does not appear to increase periodontal destruction. We think that IgG2 anti-A. actinomycetemcomitans is playing a role in limiting the extent and severity of disease in patients genetically susceptible to EOP. J Periodontol 1996;67:317-322.