Analysis of 16 cases of maxillofacial space infection caused death / 实用口腔医学杂志

16 death cases of oral and maxillofacial space infection were analyzed.10 of the 16 patients were over 60 years old,the other 6 were aged 49-57 years,most of them had systemic underlying disease,and 11 of them were with diabetes mellitus.Odontogenic infection is the leading cause of maxillofacial infections(n =13).Lethal factor in oral and maxillofacial space infection is various,and comprehensive treatment should be given.

Identification of dengue II virus-binding proteins from Aedes albopictus and Culex. Quinguefasciatus / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen DENV-2 binding proteins from Aedes albopictus and Culex. quinquefasciatus.</p><p><b>METHODS</b>The total proteins of Aedes albopictus and Culex. quinquefasciatus in different developmental stages were prepared and analyzed with SDS-12% polyacrylamide gel. After electrophoresis the proteins were transferred using Mini Trans-Blot Electrophoretic Transfer Cell (Bio-Rad ) to a nitrocellulose membrane. Virus overlay protein-binding assay (VOPBA) was carried out using anti-dengue virus 1-4 monoclonal antibody.</p><p><b>RESULTS</b>In Aedes albopictus, VOPBA detected DEN-2 binding molecules of 25 000, 35 000, and 50 000 in larvae samples, molecules of 35 000 and 50 000 in pupae samples, a 50 000 molecule in male mosquito samples, and molecules of 35 000 and 50 000 in female mosquito samples. DENV-2 binding protein of 35 000 was found in the larvae, pupae, and female mosquitoes, but not in male mosquitoes. In Culex. Quinquefasciatus, VOPBA detected a molecule of 100 000 in larvae samples, molecules of 40 000, 100 000, and around 50 000 (48 000 and 60 000) in pupae samples, and molecules of 40 000 and 100 000 in male mosquitoes and female mosquito samples.</p><p><b>CONCLUSION</b>Several proteins capable of binding DENV are found in Aedes albopictus and Culex. quinquefasciatus in different development stages. The 35 000 molecule expressed in Aedes albopictus as a putative receptor protein may be related to virus tropism in mosquito tissues.</p>

An inverted-repeat RNA construct for silencing dengue virus type 2 pre-membrane gene suppresses viral replication in BHK-21 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the anti-viral effects of a plasmid expressing an inverted-repeat RNA targeting dengue virus type-2 (DENV-2) pre-membrane (prM) gene.</p><p><b>METHOD</b>Suckling mice were inoculated with live DENV-2 in the brain. The total RNA was extracted from the brain tissue of the infected mice, and the prM gene fragments were amplified by RT-PCR and then subcloned into XhoI/EcoR I of the pcDNA3.1(+) plasmid in antisense orientation to construct the plasmid pcDNA-asprM. DENV-2 prM sequences were also subcloned into pMD18-T-vector in sense orientation to construct the plasmid pMD18-T- prM. pcDNA-irRNA was constructed by inserting in sense orientation the prM fragment isolated from pMD18-T-prM into the NheI/Kpn I of pcDNA-asprM. The plasmid pcDNA-irRNA was transfected into BHK-21 cells and the anti-viral effects were analyzed by semi-quantitative PCR and real-time PCR.</p><p><b>RESULTS</b>Transfection with the plasmid pcDNA-irRNA caused a reduction of NS3 mRNA expression level by 28% in BHK-21 cells following a 96-h challenge with DENV-2 as compared to the cells without plasmid transfection (positive control). The viral copies in pcDNA-irRNA-transfected cells was 1.44-fold lower than those in the positive control cells following a 72-h virus challenge, and the mRNA expression levels of NS1 were also significantly lower in the transfected cells at 96 h after viral challenge (P<0.05) as shown by real-time quantitative PCR.</p><p><b>CONCLUSION</b>The inverted-repeat RNA for DENV-2 prM gene silencing can suppress DENV-2 replication in BHK-21 cells, which provides a basis for developing dengue virus gene vaccine.</p>

Screening of dengue II virus-binding molecules from Aedes albopictus C6/36 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen the molecules binding dengue II virus expressed in Aedes albopictus C6/36 cells and characterize their biological functions.</p><p><b>METHODS</b>Aedes albopictus C6/36 cells were infected with dengue II virus, and the virus were collected and purified. The total and membrane proteins of C6/36 cells were extracted and analyzed using 12% SDS-polyacrylamide gel (PAGE). After electrophoresis, the proteins were transferred to a nitrocellulose membrane, and virus overlay protein-binding assay (VOPBA) was carried out using an anti-dengue virus 1-4 monoclonal antibody.</p><p><b>RESULTS</b>Two specific bands of 67 000 and 30 000 occurred after VOPBA of the proteins from the cells incubated with the virus, while the negative control group did not show these specific bands.</p><p><b>CONCLUSION</b>Two putative dengue virus receptor molecules of 67 000 and 30000 have been obtained from C6/36 cells using VOPBA, and their functional identification is in progress.</p>

Susceptibility of Aedes albopictus and Culex pipiens quinquefasciatus to infection with bat Japanese encephalitis virus isolates / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the susceptibility of Aedes albopictus and Culex pipiens quinquefasciatus to oral infection with bat Japanese encephalitis virus isolates (GD1 and HN2 strains).</p><p><b>METHODS</b>Aedes albopictus and Culex pipiens quinquefasciatus were infected orally by GD1 and HN2 strains of bat Japanese encephalitis virus. TaqMan real-time PCR was used to detect the virus and monitor the changes in the viral loads in Aedes albopictus and Culex pipiens quinquefasciatus at a 2-day interval, starting from 4 days till 20 days after the infection.</p><p><b>RESULTS</b>The infected Aedes albopictus and Culex pipiens quinquefasciatus were found positive for the Japanese encephalitis virus from day 4 to day 20. Both Aedes albopictus and Culex pipiens quinquefasciatus were susceptible to infection by GD1 and HN2 strains, but the latter showed a greater susceptibility. The HN2 strain virus appeared to have a greater virulence than the GD1 strain.</p><p><b>CONCLUSION</b>Aedes albopictus and Culex pipiens quinquefasciatus can carry GD1 and HN2 strains of bat Japanese encephalitis virus isolates.</p>

Role of estrogen-related receptor alpha in adipocyes lipolysis / 生物工程学报

Estrogen-related receptor a (ERRalpha) is a key regulator for energy metabolism and adipogenesis. However, its role in lipolysis is unknown. To study the function of ERRalpha in lipolysis, primary cultured differentiated porcine adipocytes were treated by a specific inverse agonist XCT790 or infected with adenoviral vector expressed ERRalpha for 48 h, in the absence and/or presence of specific protein kinase A (PKA) inhibitor or extracellular signal-related kinase (ERK) inhibitor. Then, we measured the triglyceride (TG) content and the glycerol release into the culture media to analysis the effect of ERRalpha on lipolysis; Further, we analyzed the expression of PPARgamma, perilipin A, p-perilipin A, HSL and ATGL with Western blotting. Here, we found that ERRalpha significantly increased adipocytes differentiation, TG accumulation and glycerol release. Separately or simultaneously block the PKA and ERK pathway do not significantly altered the effect of ERRalpha on glycerol release. ERRalpha significantly up-regulated the proteins expression of PPARgamma, perilipin A, HSL and ATGL, while the p-perilipin A protein level was not significantly changed. These findings imply that ERRalpha could increase lipolysis via up-regulating HSL and ATGL, thereby to supply more FFA as substrate for a larger turnover of cellular triglyceride pool during adipocytes differentiation.

Comparative clinical effectiveness of donepezil versus huperzine in elderly patients with mild cognitive impairment / 中华老年医学杂志

Objective To assess the effectiveness of donepezil versus huperzine in the treatment of elderly patients with mild cognitive impairment (MCI).Methods Total 122 elderly patients with MCI were divided into two groups:donepezil treatment (5.0 mg once daily) (n=71) and huperzine treatment group (0.1 mg twice daily) (n=51).All the patients were followed up for 24 weeks.Before and 12 weeks,24 weeks after drug treatment,the cognitive functions were evaluated,including MMSE,MOCA,ADAS-cog,CDR,GDS,ADL,HIS and HAMD.Results There was no significant difference in age,sex,education time and neuropsychology rating scales between the groups before drug use.As compared with the score before drug use,the donepezil group showed a significant increase in MMSE after 12-weeks (t=4.47) or 24-weeks (t=6.16) (P＜0.01),a decrease in the score of ADAS-cog after 12-weeks (t=2.33,P＜0.05) or 24-weeks( t=3.68,P＜0.05),and an increase in the score of MOCA after 24-weeks drug use (t=2.56,P＜0.05).The huperzine group showed significant improvement in MMSE after 24-weeks drug use (t=2.80,P＜0.05),but there was no difference in other time points or in the score of MOCA and ADAS-cog as compared with the score before drug use.After 24 weeks' treatment,the donepezil group had higher MMSE (t=2.01,P＜0.05) and lower ADAS-cog (t=2.09,P＜0.05) scores than the huperzine group.30 patients (total effective rate was 42.3 ％) and 9 patients (total effective rate was 17.6 ％ ) became improved in donepezil and huperzine group,respectively,with significant difference (x2 =8.26,P＜0.01 ).There were 5 cases in the donepezil group and 3 cases in the huperzine group getting slight side-effects which disappeared by continuing to take drugs or by adjusting drug taking time.Conclusions Donepezil and huperzine as the cholinesterase inhibitors are effective and safe,and the efficacy of donepezil is faster and better in treating elderly patients with MCI.

Cloning, expression of the porcine estrogen-related receptor alpha gene and its effect on lipid accumulation in mature adipocytes / 生物工程学报

Estrogen-related receptor alpha (ERR alpha) is an orphan nuclear receptor and functions as a key regulator of energy metabolism in high energy demand tissues. However, its role in white adipose tissue is largely unknown. In this study, we aim to clone the ORF sequence of pig ERR alpha with touch down-PCR, analyze the expression pattern of ERR alpha protein and its subcellular localization with Western blotting and cell immunofluorescence method respectively, and identify the effect of ERR alpha on lipid accumulation in mature porcine adipocytes with its specific inhibitor XCT790. The results showed that the ERR alpha ORF sequence is 1269 bp (GenBank Accession No. FJ446485, not published), and encode 422 amino acids. The homologies of ERR alpha nucleotide and amino acids sequences are high in different species. ERR alpha protein is highly expressed in pig white adipose tissue, kidney and heart, while remarkably lower in spleen. Cell immunofluorescence results indicated that ERR alpha protein distribute widely in adipocytes nucleus and cytoplasm. XCT790 significantly inhibited the expression of ERR alpha and lipid accumulation in porcine mature adipocyte. This study will provide new target and theoretical reference for fat deposition control.

Effects of leptin on porcine primary adiocytes lipolysis and mRNA expression of key lipolytic enzymes / 生物工程学报

Leptin, a cytokine predominantly secreted from fat tissue, plays an important role in regulating organism energy balance. Leptin can stimulate lipolysis, but the mechanism is unclear. In order to study the molecular mechanism of leptin stimulating lipolysis, we systemically studied the mRNA expression of key lipolytic enzymes. Morphological observation, Oil Red O staining and RT-PCR were used to identify pig primary adipocytes; commercial kits were used to measure the glycerol and FFA release; Semiquantitative RT-PCR was used to detect the mRNA expression of key lipolytic enzymes. The results showed that 100 nmol/L leptin up-regulated the mRNA expression of ATGL, TGH-2, HSL, MGL and LPL (P<0.01), but down-regulated the Perilipin mRNA expression (P<0.01). At the same time, leptin promoted the glycerol release in a dose dependent manner (P<0.01), but had no effect on the FFA release (P>0.05). These indicate that leptin may mainly stimulate lipolysis in pig primary adipocytes by up-regulating the expression of ATGL, MGL, LPL and down-regulating the expression of Perilipin. The unchanged FFA release may be resulted from Leptin promoting UCPs mRNA expression and increasing FFA expenditure.

CLONING AND SEQUENCE ANALYSIS OF SSU rRNA GENE OF CUTANEOUS LEISHMANIASIS PATHOGEN FROM XINJIANG OF CHINA / 中国寄生虫学与寄生虫病杂志

Objective] By sequencing of SSU rRNA gene cloning from Xinjiang cutaneous leishmaniasis pathogen (XJCLP) to provide evidence for identification of the pathogen. [Methods] By PCR assay with primers R222 and R333, the specific fragment had been produced from SSU rRNA gene of XJCLP , L infantum, L tropica and cloned into pGEM ○[KG-6/7]R T Easy vector .The clones were sequenced by the Sanger dideoxy mediated chain termination method, analysis of SSU rRNA gene sequences from XJCLP, L tropica, L infantum with DNASIS. [Results] Sequence analysis showed that the specific fragment of SSU rRNAgene from XJCLP, L infantum,L tropica , were all 394 bp in length. There were 391 bases identical and three point mutations between the sequences of XJCLP and L tropica , the similarity being 99 2%; 390 bases identical and three point mutations and one insertion /deletion between the sequences of XJCLP and L infantum , the similarity being 99 0%. One insertion/deletion between the sequences of L tropica and L infantum , the similarity being 99 7%. The primary and secondary structures of SSU rRNA gene from XJCLP differed from those of L infantum and L tropica .A retrieval from GenBank confirmed that these 394 bp sequence are new gene sequences. [Conclusion]The primary and secondary structures of SSU rRNA gene from XJCLP, L infantum , L tropica were different. 394 bp sequence from SSU rRNA gene of XJCLP is a new gene sequence.

Cloning and Identification of the cDNA and Genomic DNA Sequences of the Defensin Gene of Anopheles sinensis / 中国寄生虫学与寄生虫病杂志

Objective To clone and identify the cDNA sequence and genomic DNA sequence of Anopheles sinensis defensin gene. Methods Referring to the published defensin gene sequences of Aedes aegypti and Anopheles gambiae, pairs of primers were designed to amplify the cDNA sequence and genomic DNA sequence of Anopheles sinensis defensin gene with template of Anopheles sinensis total RNA by RT-PCR and genomic DNA by nested PCR, respectively. These amplified fractions were cloned and sequenced, and were further identified and analyzed by relevant bioinformatics softwares. Results Complete genomic DNA sequence was cloned, including 5′ and 3′ UTR fraction and two exons separated by a 85 bp intron. Besides, the whole cDNA sequence of Anopheles sinensis defensin gene with 324 bp was also cloned, and its ORF encoded 107 amino acids. The mature peptide had 40 amino acids residues. Conclusion The whole cDNA sequence and complete genomic DNA sequence of the defensin gene of Anopheles sinensis have been cloned and identified for the first time.