Study on rAd5F35-SIVenvT vaccine in combination with rMVA-SIVenvT vaccine in mice / 中华微生物学和免疫学杂志

Objective:To analyze the immune response in mice after immunization with vaccine of rAd5F35-SIVenvT in combination with rMVA-SIVenvT to evaluate the efficacy of different immunization strategies.Methods:Two recombinant viruses were identified in vitro by PCR and Western blot. The BALB/c mice were immunized with homologous and heterologous immune strategies. The numbers of splenic lymphocytes secreting IFN-γ were measured by ELISPOT assay, meanwhile SIV gp120 antibody titer were measured by ELISA assay. Results:SIVenvT protein was expressed effectively by rAd5F35-SIVenvT and rMVA-SIVenvT in HEK293 cells. The specific immune response reached its peak at 4-week post first immunization, then decreased. SIV Env specific cellular immune response and SIV gp120 specific antibody could be detected at 4-16 weeks post first immunization. The specific cellular response was significant stronger in heterologous immunization group than homologous group at 4 week and 16 week. Furthermore, heterologous immunization induced significant higher titer of SIV gp120 antibody at 4 week than homologous group.Conclusions:Specific immune response induced by rAd5F35-SIVenvT in combination with rMVA-SIVenvT was stronger than homologous vector immunization. The results provided references for further study in nonhuman primates.

Serological and molecular techniques applied for identification of Plasmodium spp. in blood samples from nonhuman primates / Técnicas sorológicas e moleculares aplicadas na identificação de Plasmodium spp. em amostras de primatas não humanos

Abstract The aim of this study was to identify Plasmodium spp. in blood samples from nonhuman primates (NHPs) in the state of Maranhão, using classical and alternative techniques for examination of human malaria. A total of 161 blood samples from NHPs were analyzed: 141 from captive animals at a Wildlife Screening Center (CETAS) and 20 from free-living animals in a private reserve. The techniques used were microscopy, rapid diagnostic test (RDT), Indirect fluorescent antibody test (IFAT) and molecular techniques (semi-nested PCR, quantitative real-time PCR and LAMP). Two serological methods (dot-ELISA and indirect ELISA) were also standardized with rhoptry protein-soluble antigen of P. falciparum and P. berghei. Trophozoite forms of Plasmodium sp. were identified on slides from five different animals. No samples were positive through RDT and LAMP. Four samples were seropositive for P. malariae through IFAT. The samples showed low reactivity to ELISA. Plasmodium sp. was detected in 34.16% (55/161) of the samples using qPCR based on the 18S rRNA gene. After sequencing, two samples showed 100% identityl to P. malariae, one showed 97% identity to Plasmodium sp. ZOOBH and one showed 99% identity to P. falciparum . PCR was shown to be the most sensitive technique for diagnosing Plasmodium in NHP samples.

Resumo Neste estudo objetivamos identificar Plasmodium spp. em amostras sangue de primatas não humanos (PNH) do estado do Maranhão, utilizando técnicas clássicas e alternativas para o exame da malária humana. Foram analisadas 161 amostras de sangue de PNH, sendo 141 de CETAS (cativeiro) e 20 de reserva particular (vida livre), utilizando microscopia, teste de diagnóstico rápido (RDT), imunofluorescência indireta (IFI) e técnicas moleculares (semi-nested PCR, PCR em tempo real quantitativo e LAMP). Dois métodos sorológicos (dot-ELISA e ELISA indireto) também foram padronizados com antígenos solúveis de roptrias de P. falciparum e P. berghei. Formas trofozoíticas de Plasmodium sp. foram identificadas em lâminas de cinco animais diferentes. Nenhuma amostra foi positiva em TDR e LAMP. Quatro amostras foram soropositivas para P. malariae na IFI. Os soros de PNH mostraram baixa reatividade pelo ELISA indireto. Plasmodium sp. foi detectado em 34,16% (55/161) das amostras utilizando a qPCR baseada no gene 18S rRNA. No sequenciamento, duas amostras mostraram identidade com P. malariae (100%), uma com Plasmodium sp. ZOOBH (97%) e uma com P. falciparum (99%). A PCR mostrou ser a técnica mais sensível para diagnósticos de Plasmodium em amostras de PNH.

Changes in the expression of CD169 on peripheral blood CD14 +monocytes of rhesus monkeys after SIV infection / 中国比较医学杂志

Objective To investigate the changes of CD169 expression on the surface of peripheral blood monocytes and different subsets of monocytes in normal rhesus monkeys after SIVmac239 infection and the possible reasons.Methods Normal rhesus monkeys were infected with SIVmac239 through intravenous injection, and changes in the percentage of peripheral blood monocytes and the expression of CD169 before and after SIVmac239 infection were detected by flow cytometry. The peripheral blood CD14 +monocytes of normal rhesus monkeys sorted by flow cytometry were directly infected by SIVmac239 and stimulated by different cytokines,and changes in the expression of CD169 on the cell surface and the cytokine IFN-α were detected by flow cytometry. Results After SIVmac239 infection, the percentage of CD14 +monocytes of the normal rhesus monkeys was decreased and the expression of CD169 on their surface was increased. Meanwhile,the expression of CD169 on the surface of different subsets of peripheral blood monocytes was significantly increased,and the expression of CD169 on the CD14 +CD16 + +monocytes was increased more obviously. CD169 was not expressed on the surface of peripheral blood CD14 +monocytes of the normal rhesus monkeys after stimulated by the cytokines M-CSF, IL-4 and IL-13. However, CD169 was highly expressed after the monocytes were stimulated by the cytokine IFN-α. The expression of CD169 on the surface of CD14 +monocytes and the intracellular cytokine IFN-α was not significantly changed after the monocytes were directly infected with SIVmac239. Conclusions SIVmac239 infection can lead to the increase of CD169 expression on the surface of peripheral blood monocytes in rhesus monkeys. Its expression is not associated with the direct infection of virus,but is related to the cytokine IFN-α secreted by other cells of the monkeys in vivo.

Establishment and application of a new immune-comb assay for detection of serum antibody against simian immunodeficiency virus / 中国实验动物学报

Objective SIV30 protein of simian immunodeficiency virus(SIV)was prepared by genetic engineering technique as an antigen diagnostic reagent, to establish an immune comb method for the specific detection of anti SIV IgG in monkey serum. Methods Recombinant expression plasmid of SIV SIV30 gene was constructed by prokaryotic expression vector pGEX-4T-1, and expressed in the competent BL21 cells. The recombinant protein was purified as a diagnostic antigen, and a standardized procedure for the detection of immune comb was established and applied for clinical detection. Results The optimum coating amount of antigen was 0.02 mg/mL. The prepared IC was able to specifically detect the positive serum of SIV. There was no cross reaction between the sera of other viruses. It showed a high specificity of the detection method. Sensitivity analysis showed that the SIV30 protein was able to detect 1:400 times diluted SIV positive sera. The result of stability and repeatability test(the same sample was repeated 3 times) showed that the coefficient of variation(CV)was less than 10%. The serum samples of 10 suspicious monkeys were detected by this method, showing a consistent rate of comb method and ELISA test result of 100%, Kappa =1.000. Conclusions SIV30 protein is expressed in prokaryotic cells. The immune comb is prepared,and is successfullyl applied in clinical examination. It shows that the method has a high sensitivity, strong specificity, good reproducibility and practicability.

Detection of Plasmodium in faeces of the New World primate Alouatta clamitans

Abstract Plasmodium falciparum and Plasmodium vivax have evolved with host switches between non-human primates (NHPs) and humans. Studies on the infection dynamics of Plasmodium species in NHPs will improve our understanding of the evolution of these parasites; however, such studies are hampered by the difficulty of handling animals in the field. The aim of this study was to detect genomic DNA of Plasmodium species from the faeces of New World monkeys. Faecal samples from 23 Alouatta clamitans from the Centre for Biological Research of Indaial (Santa Catarina, Brazil) were collected. Extracted DNA from faecal samples was used for molecular diagnosis of malaria by nested polymerase chain reaction. One natural infection with Plasmodium simium was identified by amplification of DNA extracted from the faeces of A. clamitans. Extracted DNA from a captive NHP was also used for parasite genotyping. The detection limit of the technique was evaluated in vitro using an artificial mixture of cultured P. falciparum in NHP faeces and determined to be 6.5 parasites/µL. Faecal samples of New World primates can be used to detect malaria infections in field surveys and also to monitor the genetic variability of parasites and dynamics of infection.

Establishment of a real-time fluorescent quantitative PCR for the rapid detection of viral reservoir in the peripheral blood of SIV-infected rhesus monkeys / 中华微生物学和免疫学杂志

Objective To establish a TaqMan probe-based real-time fluorescent quantitative PCR ( real-time PCR) for the quantitative and rapid detection of viral reservoir in peripheral blood mononuclear cells (PBMCs) isolated from rhesus monkeys with simian immunodeficiency virus (SIV) infection and to evaluate its preliminary application. Methods A pair of primers and one TaqMan probe were designed ac-cording to the conserved sequence of SIVmac239 strain for real-time PCR amplification. A length of 2 090 bp of nucleotide fragment was digested from the plasmid p239SpSp5 containing 5′-end long segments of SIV-mac239 strain by restriction enzymes EcoRⅠand SpeⅠ. The standards used for quantitative detection of SIV DNA in peripheral blood samples were prepared by a 10-fold serial dilution and used for graphing the stand-ard curve. The numbers of SIV DNA ( copies per 106 PBMCs) in rhesus monkeys during acute and chronic phases of SIVmac239 infection were determined and the virological characteristics of SIV DNA at different phages of infection were analyzed. Results A linear positive correlation between cycle threshold ( Ct) val-ues and concentrations (10 copies/μl to 109 copies/μl) of the standards was found. High levels of SIV DNA were monitored in SIV-infected monkeys 14 to 22 days after acute infection. The levels of SIV DNA in the acute phase of infection were about 1 to 2 logs higher than those in the chronic phase of infection. The num-bers of SIV DNA ( copies per 106 PBMCs) were 1 log lower than the SIV viral load in peripheral blood of the same monkey. The ratios of SIV DNA load to SIV RNA load ( DNA/RNA) in chronic phase of infection were higher than those in the acute phase. Conclusion The established TaqMan probe-based real-time fluorescent quantitative PCR was a highly sensitive and specific assay for the detection of SIV DNA with an advantage of wide linear range. It could be used for the quantitative evaluation of latent reservoirs of SIV.

Unilateral simian crease in a 3 years old child - A case report.

Unilateral simian crease (USC) is a single transverse crease that extends from across the palm of one of the hands. The presence of a single transverse palmar crease or the simian crease (SC) can be seen in normal individuals. The significance of USC lies in the fact that it can also be associated with abnormal medical conditions. Literature review indicates that there is strong coincidence with the presence of a SC and presence of genetic or chromosomal abnormalities in these subset of patients. USC is seen in 10% of the population. In this case report, a three year old child was detected to have a SC, especially an USC in one of his palms.

Discriminant Analysis of Platform Parameters of Rhesus at Different Stages of SIV/SAIDS / 广州中医药大学学报

Objective To predict the disease progression risks of healthy rhesus ( normal) and rhesus infected with simian immunodeficiency virus ( SIV) in the stages of long-term nonprogressor ( LTNP) , normal progressor ( NP) , rapid progressor ( RP) by discriminant analysis. Methods Five-year observation was carried out in SIV infected rhesus model without any intervention. The SIV infected rhesus model at the stages of LTNP, NP, RP were selected, 10 in each group, and T lymphocyte subsets and serum parameters for spleen-deficiency syndrome and kidney-deficiency syndrome in SIV infected rhesus were compared with 5 healthy monkey having the same survival time. The influence factors of different types of disease progression were screened from T cell subsets and Chinese medical syndrome indexes, and then the discriminant equation was established to predict the risks. Results White blood cell ( WBC) count and lymphocyte ( LYM) ratio were enrolled into the discriminant equation before infection, and T4 level and Log10RNA of set point were enrolled into the discriminant equation in the platform period. The test results for the uniform rate of the established discriminant function showed that the total coincidence rate of theoretic distinguish to the actual data was 57.1% , 91.2%respectively before infection and in the platform period. Conclusion The pre-infection WBC count and LYM ratio can be used as a reference for the evaluation of different types of disease progresson, and Log10RNA and T4 level at platform phase can be used as the predicting factors of different types of disease progression risk prediction.

Malaria in Brazil: what happens outside the Amazonian endemic region

Brazil, a country of continental proportions, presents three profiles of malaria transmission. The first and most important numerically, occurs inside the Amazon. The Amazon accounts for approximately 60% of the nation’s territory and approximately 13% of the Brazilian population. This region hosts 99.5% of the nation’s malaria cases, which are predominantly caused by Plasmodium vivax (i.e., 82% of cases in 2013). The second involves imported malaria, which corresponds to malaria cases acquired outside the region where the individuals live or the diagnosis was made. These cases are imported from endemic regions of Brazil (i.e., the Amazon) or from other countries in South and Central America, Africa and Asia. Imported malaria comprised 89% of the cases found outside the area of active transmission in Brazil in 2013. These cases highlight an important question with respect to both therapeutic and epidemiological issues because patients, especially those with falciparum malaria, arriving in a region where the health professionals may not have experience with the clinical manifestations of malaria and its diagnosis could suffer dramatic consequences associated with a potential delay in treatment. Additionally, because the Anopheles vectors exist in most of the country, even a single case of malaria, if not diagnosed and treated immediately, may result in introduced cases, causing outbreaks and even introducing or reintroducing the disease to a non-endemic, receptive region. Cases introduced outside the Amazon usually occur in areas in which malaria was formerly endemic and are transmitted by competent vectors belonging to the subgenus Nyssorhynchus (i.e., Anopheles darlingi, Anopheles aquasalis and species of the Albitarsis complex). The third type of transmission accounts for only 0.05% of all cases and is caused by autochthonous malaria in the Atlantic Forest, located primarily along the southeastern Atlantic Coast. They are caused by parasites that seem to be (or to be very close to) P. vivax and, in a less extent, by Plasmodium malariae and it is transmitted by the bromeliad mosquito Anopheles (Kerteszia) cruzii. This paper deals mainly with the two profiles of malaria found outside the Amazon: the imported and ensuing introduced cases and the autochthonous cases. We also provide an update regarding the situation in Brazil and the Brazilian endemic Amazon.

Plasmodium simium/Plasmodium vivax infections in southern brown howler monkeys from the Atlantic Forest

Blood infection by the simian parasite, Plasmodium simium, was identified in captive (n = 45, 4.4%) and in wild Alouatta clamitans monkeys (n = 20, 35%) from the Atlantic Forest of southern Brazil. A single malaria infection was symptomatic and the monkey presented clinical and haematological alterations. A high frequency of Plasmodium vivax-specific antibodies was detected among these monkeys, with 87% of the monkeys testing positive against P. vivax antigens. These findings highlight the possibility of malaria as a zoonosis in the remaining Atlantic Forest and its impact on the epidemiology of the disease.

Comparative analysis of Gp120 sequences of enterotropic and neurotropic SIV strains / 中国比较医学杂志

Objective To study the mutations of Env sequence of SIVmac239 after infection of Chinese rhesus monkeys, and compare the differences and characteristics of Gp120 sequences of enterotropic and neurotropic SIV strains. Methods Six strains of simian immunodeficiency virus were analyzed in this study: four separated from peripheral blood mononuclear cells of SIVmac239-infected monkeys and two neurotropic SIVmac251 strains.Isolated and cultured monoclonal virus was obtained by limiting dilution assay.Gp120 sequences were amplified after the RNA extraction and phylogenetic analysis was processed thereafter.So did the Gp120 amino acid sequence and N-glycosylation sites analysis of the enterotropic and neurotropic strains.Results SIVmac239 had different mutations in four rhesus monkeys.The diversity in amino acid sequences of the enterotropic and neurotropic strains concentrated in the V1 and V4 regions of Gp120.The enterotropic strains had an addition of glycosylation site in V4 but the glycosylation site changes of neurotropic strains were located in the conservative regions of C1, C2 and C3.Conclusions The addition of one glycosylation site in V4 region of GP120 and loss of one glycosylation site in C1 region are associated with enhanced enterotropism and neurotropism.The differences between the enterotropic and neurotropic strains are not dipicted in Gp120 V3 region which is closely related with the tropism of strains.

Well-Differentiated Papillary Mesothelioma of the Tunica Vaginalis: A Case Study and Review of the Literature

Well-differentiated papillary mesothelioma is an uncommon tumor of the testes that usually presents as a hydrocele. Here, we present the case of one patient who did not have a history of asbestos exposure. The tumor was localized in the tunica vaginalis and was composed of three pedunculated masses macroscopically. Microscopically, branching papillary structures with focal coagulative necrosis were present. In addition to immunohistochemistry, simian virus 40 DNA was also tested by polymerase chain reaction. This report presents one case of this rare entity, its clinical and macroscopic features, and follow-up results.

Effect of repeated rectal exposure of low-dose simian immunodeficiency virus on the systemic cellular immunity in monkeys / 中国比较医学杂志

Objective To study the effect of repeated rectal exposure of low -dose simian immunodeficiency virus on the systemic cellular immunity in monkeys .Methods Eight 3-to 4-year old rhesus macaques ( Macaca mulatta) (male:female 1:1) were used in this study.The monkeys were inoculated with 10 TCID50 SIVmac239 virus through rectum twice a week for consecutive 6 weeks to establish a multiple rectal exposure model of SIVmac 239 virus infection.Then, plasma viral load, CD4+ T cell count, T cell subsets and IFN-γsecretion of the experiment monkeys were determined . Results Low-dose SIVmac239 virus induced some changes in the immune system through the rectal mucosa , but didn’t induce typical infection.Repeated rectal mucosal low-dose virus exposure can activate the cellular immune system . Conclusions This study defines the effect of repeated low -dose simian immunodeficiency virus exposure on the systemic cellular immunity, and provided basic information for HIV-1 vaccine research.

No Detection of Simian Virus 40 in Malignant Mesothelioma in Korea

BACKGROUND: Simian virus 40 (SV40), a polyomavirus, was discovered as a contaminant of a human polio vaccine in the 1960s. It is known that malignant mesothelioma (MM) is associated with SV40, and that the virus works as a cofactor to the carcinogenetic effects of asbestos. However, the reports about the correlation between SV40 and MM have not been consistent. The purpose of this study is to identify SV40 in MM tissue in Korea through detection of SV40 protein and DNA. METHODS: We analyzed 62 cases of available paraffin-blocks enrolled through the Korean Malignant Mesothelioma Surveillance System and performed immunohistochemistry for SV40 protein and real-time polymerase chain reaction (PCR) for SV40 DNA. RESULTS: Of 62 total cases, 40 had disease involving the pleura (64.5%), and 29 (46.8%) were found to be of the epithelioid subtype. Immunostaining demonstrated that all examined tissues were negative for SV40 protein. Sufficient DNA was extracted for real-time PCR analysis from 36 cases. Quantitative PCR of these samples showed no increase in SV40 transcript compared to the negative controls. CONCLUSIONS: SV40 is not associated with the development of MM in Korea.

The Comparison of Genetic Variation in the Envelope Protein Between Various Immunodeficiency Viruses and Equine Infectious Anemia Virus / 中国病毒学·英文版

The envelope protein(Env) of lentiviruses such as HIV,SIV,FIV and EIAV is larger than that of other retroviruses.The Chinese EIAV attenuated vaccine is based on Env and has helped to successfully control this virus,demonstrating that envelope is crucial for vaccine.We compared Env variation of the four kinds of lentiviruses.Phylogenetic analysis showed that the evolutionary relationship of Env between HIV and SIV was the closest and they appeared to descend from a common ancestor,and the relationship of HIV and EIAV was the furthest.EIAV had the shortest Env length and the least number of potential N-linked glycosylation sites(PNGS) as well as glycosylation density compared to various immunodeficiency viruses.However,HIV had the longest Env length and the most PNGS.Moreover,the alignment of HIV and SIV showed that PNGS were primarily distributed within extracellular membrane protein gp120 rather than transmembrane gp41.It implies that the size difference among these viruses is associated with a lentivirus specific function and also the diversity of env.There are low levels of modification of glycosylation sites of Env and selection of optimal protective epitopes might be useful for development of an effective vaccine against HIV/AIDS.

Comparison of bio-medical parameters in SIV infected Chinese rhesus monkeys with diverse progression correlated to the pathogenesis of simian AIDS / 中华微生物学和免疫学杂志

Objective To compare the bio-medical parameters in SIV infected Chinese rhesus monkeys with diverse disease progression,by which the pathogenesis of simian AIDS were to be investigated.Methods Sixteen Chinese rhesus monkeys were inoculated intravenously with SIVmac239 and followed-up for 18 months.Based on their progression patterns and plasma viral loads,animals were divided into 3 groups,including 1 rapid progressor( RP),13 normal progressors(NP),and 2 elite controllor(EC).Their parameters of haematology,virology,immunology and pathology were examined and compared. Results Compared with other animals,RM449(RP) showed higher viral load,unresponsive humoral immunity,and higher level of auto-antibodies against lymph node,thymus,and spleen.Additionally,its effector memory CD4 count was lower,with the transformation progress being blocked-like from naive/central memory subsets to effector memory subset,as the flow-cytometry assay showed.Notable decrease in its peripheral B cell was also observed,especially to the sub-population of tissue-like memory B cells and activated memory B cells.Pathological examination showed the depletion of lymphoid tissue,atrophy of spleen and loss of thymus.Moreover,most of these parameters of RM450 and RM453 (EC) changed opposite to that of RP.Conclusion The hallmarks of RM449 were higher viraemia and lower SIV specific IgG level,which may due to the disturbance of T cells and B cells development and differentiation.Moreover,destructions of organs of the immune system may contribute to the disturbance.Our study suggest that the change of micro-environments of thymus induced by SIV infection,which is necessary in T cell and B cell development and differentiation,may contribute at least partially to the AIDS pathogenesis.

Prevalences of simian, Sydney and Suwon creases and their association with each other, body sides, handedness, sex and anomalies/diseases/syndromes in a population of Central India / Prevalencias de liegues simiesco, de Sydney y de Suwon y su asociación entre ellos, lados del cuerpo, dominancia manual, sexo y anomalías/enfermedades/síndromes en una población de la India Central

The Simian and Sydney creases are variant palmar creases which have now drawn medical attention as their presence correlates strongly with numerous chromosomal anomalies and diseases. Works on these creases have been done on several human communities, racial and ethnic but no specific study is focussed out on the Indian populations. In this study 1000 Central Indian subjects (500 males and 500 females) aged between 5 to 70 years were randomly selected and examined by the authors. We found the prevalence of Simian, Sydney and Suwon creases in studied population of Central India is 14.4 percent, 3.6 percent and 2.4 percent respectively, which is a little higher than the figures for Asians and Caucasians claimed by earlier studies. There is no any association between these creases. The Simian crease is predominantly unilateral right side and associated with handedness. It is more common in males. The Sydney crease does not have unilateral or bilateral predominance and association with handedness but it is more common in females. The Suwon crease does not have unilateral or bilateral predominance and association with handedness but it is more common in males. There is no association of these creases with anomalies/diseases/syndromes. Conclusively this work draws attention to the importance of undertaking parallel investigations on every socio-cultural human group or population wherever possible.

Los pliegues simiescos y de Sydney, son pliegues palmares variantes que en la actualidad han llamado la atención médica, ya que su presencia se relaciona estrechamente con numerosas anomalías cromosómicas y enfermedades. Investigaciones sobre estos pliegues se han realizado en diferentes comunidades humanas, raciales y étnicas, pero ningún estudio específico se centra sobre las poblaciones indígenas. En este estudio 1000 sujetos de la India central (500 hombres y mujeres 500) entre 5 y 70 años fueron seleccionados al azar y examinados. Se encontró que la prevalencia de los pliegues Simiescos, de Sydney y de Suwon de la población estudiada es de 14,4 por ciento, 3,6 por ciento y 2,4 por ciento respectivamente, mayor que los porcentajes en asiáticos y caucásicos. No hay ninguna asociación entre estos pliegues. El pliegue simiesco es predominantemente unilateral derecho y se asocia con dominancia manual. Es más común en los hombres. El pliegue de Sydney no tiene predominio unilateral o bilateral, ni asociación con dominancia manual, pero es más común en las mujeres. El pliegue de Suwon no tiene predominio unilateral o bilateral, ni asociación con dominancia manual, pero es más común en los hombres. No existe ninguna asociación de estos pliegues con anomalías/enfermedades/síndromes. Se concluye la importancia de realizar investigaciones paralelas en todos los grupos humanos o socio-culturales de la población siempre que sea posible.

SIVmac 239 infection in Rhesus monkeys: histopathologic changes / 临床与实验病理学杂志

Purpose To observe the histopathologic changes of acquired immure deficiency syndrome (AIDS) in a Chinese Rhesus monkeys model and the pathogenesis that initiated the changes.Methods Chinese Rhesus monkeys were sacrificed after being inoculated SIVmac239 by Ⅳ(n=2)for four months.Autopsy was carried out by pathologic routine method.The lymph nodes, heart, liver, spleen, lung, kidney, digestive tract and other tissues were selected, the tissues fixed with 10% neutral formalin, and the pathologic sections were prepared by HE staining and immunohistochemical staining and special staining after paraffin imbedding.Results The main histopathological changes appeared in the immune system in different organs. The lymph nodes began to display the complex changes in a short period of time infected by the virus, including proliferation of lymphoid follicles, atrophy, or both; some lymphoid follicles of lymph nodes had few lymphocytes, with fibrous hyperplasia and immune complex (IC) deposition, displaying a burning down phenomenon.Splenomegaly and blood vessel and its endothelial cell proliferation in splenic corpuscles were noted with the immune complex deposition. Other parts of the mucosa-associated lymphoid tissue had different degrees of hyperplasia, or atrophy.Conclusion Histopathologic changes in Chinese Rhesus monkeys infected by SIVmac239 strain are very similar to human AIDS, which suggests that the model is a useful tool for the prevention and treatment study of AIDS.

Analysis of 30 cases with diabetic foot treated mainly by Simian Yongan Decoction anti Xuefu Zhuyu Decoction / 国际中医中药杂志

Objective To observe the efficacy of the treatment of diabetic foot mainly by Simian Yongan Decoction and Xuefu Zhuyu Decoction. Methods 60 patients were randomly recruited into a control group and a treatment group. The control group was treated by conventional western medicine. The treatment group was treated with Simian Yongan Decoction and Xuefu Zhuyu Decoction on the basis of western medicine. Results The total effective rate was 86.6% and 66.6% in the treatment group and the control group respectively, showing significant difference (P<0.05) . Conclusion The efficacy of Simian Yongan Decoction and Xuefu Zhuyu Decoction combined with westeru medicine was better than exclusive western medicine in treating diabetic foot.

Immortalization of Human Precartilaginous Stem Cells by Transfecting SV40Tag / 华中科技大学学报（医学）（英德文版）

Immortalized human precartilaginous stem cells (IPSCs) were established to provide stable cell resource for the study of the molecular mechanism of gene targeting on the differentiation of PSCs.Plasmid pCMVSV40T/PUR containing simian virus 40 large T antigen gene (SV40Tag) was transfected into human PSCs by using lipofectin transfection.Colonies were isolated by puromycin selection and expanded by multiple passages,lmmunohistochemistry,RT-PCR and Southern blotting were used to identify the transfected cells and to detect the expression and integration of SV40Tag in expanded cell lines.The positive colonies were isolated and subcultured,designated immortalized precartilaginous stem cells (IPSCs),which were confirmed as fibroblast growth factor receptor-3 (FGFR-3) positive cells by immunohistochemistry and RT-PCR.SV40Tag cDNA was found in cultured IPSCs of passage 8 by Southern blotting,and the expressions of SV40Tag mRNA and protein were confirmed by RT-PCR.These findings suggested that IPSCs strain with SV40Tag was constructed successfully.