The viral genome editing technologies have been shown to largely contribute to the stud-ies of viral gene functions and to the treatment of infectious diseases. With the development of genetic engi-neering technologies, a variety of viral genome mutation technologies have been established. The homologous recombination is traditionally used to edit the viral genome, but the low frequency of homologous recombina-tion limits its application. Large genome DNA viruses including herpes simplex virus 1 can be edited by bac-terial artificial chromosome system. However, the cloning of viral genome to bacterial artificial chromosome plasmid is both laborious and time-consuming, and parts of the plasmid genes or drug selection markers may remain in the genome of virus and then affect the function of virus. Fortunately, the CRISPR-cas9 system which was discovered in 2013 can be used to edit the viral genome efficiently and accurately. Furthermore, the experiment is simple and will not leave any trail of the viral genome when the genetic mutation is per-formed. Here, we briefly review the application of homologous recombination, bacterial artificial chromosome system and CRISPR-cas9 system in researches on herpes simplex virus 1.

The molecular modifications of Herpes Simplex Virus Type Ⅰ (HSV-1) proteins represented by acetylation and phosphorylation are essential to its biological functions.The cellular chromatin-remodeling/assembly is involved in HSV-1 associated gene transcriptional regulation in human cells harboring HSV-1 lytic or latent infections.Further investigation on these biological events would provide a better understanding of the mechanisms of HSV- 1 viral gene transcriptional regulation.

Herpes simplex virus type 1 (HSV-1), as a pathogen usually leading to herpes labialis, has been found to cause more cases of genital herpes and be associated with the occurrence of neurological diseases such as Alzheimer′s disease (AD). AD is a chronic and progressive degenerative disease of the central nervous system, the pathogenesis of which has not been fully elucidated, and currently there is a lack of drugs that can effectively treat the disease. The hypothesis that HSV-1 infection is involved in the pathogenesis of AD has been discussed for a long time, but there is no definite evidence to prove it. This article briefly reviewed the research progress in the pathogenesis of AD caused by HSV-1.

Nuclear proteins often form punctiform structures, but the precise mechanism for this process is unknown. As a preliminary study, we investigated the aggregation of an HSV-1 immediate-early protein,infected-cell protein 22 (ICP22), in the nucleus by observing the localization of ICP22-EGFP fusion protein.Results showed that, in high-level expression conditions, ICP22-EGFP gradually concentrates in the nucleus,persists throughout the cell cycle without disaggregation even in the cell division phase, and is finally distributed to daughter cells. We subsequently constructed a mammalian cell expression system, which had tetracyclinedependent transcriptional regulators. Consequently, the location of ICP22-EGFP in the nucleus changed with distinct induction conditions. This suggests that the cellular location of ICP22 is also influenced by promoter regulation, in addition to its own structure. Our findings provide new clues for the investigation of transcriptional regulation of viral genes. In addition, the non-protease reporter system we constructed could be utilized to evaluate the role of internal ribosome entry sites (IRES) on transcriptional regulation.

The three immediate-early proteins of HSV-1, ICPO, ICP22, and ICP27, have specific and pivotal functions in transcriptional activation and inhibition, multiple regulatory and control processes of viral genes. In this paper, the expression and localization of these three proteins were studied in neuroblastoma cells using biochemical assays, and their possible and potential interactive functions are discussed. The data show that the three proteins are localized in different structures, specifically in the PML-NB-associated structure, which is a specific nuclear structure composed of many protein molecules and bound tightly to the nuclear matrix in neuroblastoma cells. The results suggest that the activating and suppressive functions of ICPs are mostly dependent on their transcriptional and regulatory roles, including the PML-NB-associated structure.

Objective To study the diagnosis and treatment of malignant renal angiomyolipoma with metastases. Methods Three cases of advanced malignant renal angiomyolipoma with metasta-ses were retrospectively reviewed. Case 1 was a 55-year-old woman presenting with recurrent low-grade fever and aching pain in left flank. Ultrasound showed solid mass in the left kidney. Left radical nephrectomy was then performed. The right pulmonary lobectomy of the inferior lobe and wedge ex-section of superior lobe was performed 7 years later because of multiple pulmonary metastases. Case 2 was a 37-year-old woman. Left nephrectomy was performed because Ultrasound and CT showed left kidney solid mass. Six years later, multi-site metastases were found in liver and retroperitoneum and mestastasis tumors were resected. At 10 years after the primary diagnosis, CT showed multi-metasta-ses in liver and retroperitoneum. The retroperitoneal masses were resected and liver lesions were trea-ted by radiofrequency ablation. Case 3 was a 34-year-old man presenting with swelling pain in right flank. CT scan showed a lesion in the right kidney and right radical nephrectomy was performed. Four months after the surgery, MRI revealed multiple liver and retroperitoneal nodules. All the 3 cases had not been diagnosed with tuberous sclerosis and did not accept chemotherapy. Results The cut sur-face of the lesions was red-brown and yellow and the texture was tender. Under microscopic examina-tion, the tumors of case 1 and case 3 were composed of sheets or nests of large polygonal epithelioid cells. It revealed that occasionally clear cytoplasm with abundant eosinophilic, prominent nucleoli, and multinucleated and markedly pleomorphic form. Necrosis was presented as well. Large areas of case 2 tumor were made up of spindle smooth-muscle cells, adipose tissue, thick-wall blood vessels and some areas merged with a proliferation of epithelioid which was consistent with typical angiomyolipoma. Im-munohistochemical study showed that the epithelioid cells and spindle smooth-muscle cells were posi-tive for VM, HMB45, Melan-A and negative for S100, CK. Case 1 and case 3 were diagnosed with malignant epithelioid angiomyolipoma, while case 2 was diagnosed with malignant classic angiomyoli-poma and epithelioid in part of the tumor. Case 1 was well alive. Case 2 was alive with tumor 12 years after the diagnosis. And case 3 was missed in the follow-up 3 months after metastasis resection. Conclusions Malignant renal angiomyolipoma is a rare disease. The diagnosis depends on histopatho-logic, immunohistochemieal study and clinical follow-up. Radical resection of the primary, recurrent and metastatic tumors is the main therapy. It needs more research to clarify if metastasis has any effect on prognosis.

HSV-1 infection-mediated regulation of mRNA translation in host cells is a systematic and complicated process. Investigation of the details of this mechanism will facilitate understanding of biological variations in the viral replication process and host cells. In this study, a comparative proteomics technology platform was applied by two-dimension electrophoresis of HSV-1 infected normal human L-02 cell and control cell lysates. The observed protein spots were analyzed qualitatively and quantitatively by the PDQuest software package. A number of the different observed protein spots closely associated with cellular protein synthesis were identified by matrix-assisted laser-desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS). The expression levels of the RPLP1 protein, which is required for mRNA translation, and KHSRP protein, which is involved in rapid decay of mRNA, were up-regulated, whereas the expression level of RNP H2, which is involved in positive regulation on the mRNA splicing process, was down-regulated. All of these results suggest that HSV-1 infection can influence cellular protein synthesis via modulation of cellular regulatory proteins involved in RNA splicing, translation and decay, resulting in optimisation of viral protein synthesis when cellular protein synthesis is shut off. Although there is need for further investigations regarding the detailed mechanisms of cellular protein control, our studies provide new insight into the targeting of varied virus signaling pathways involved in host cellular protein synthesis.

The Sindbis-like virus was first discovered in China in 1986. Its complete genomic sequence consists of more than 11 000 bp encoding more than 3 700 amino acids. It contains a 5' non-transcriptional region (5'-NTR) in a non-structural region, four non-structural proteins (nsP1, nsP2, nsP3, nsP4) regions, capsids in conserved and non-conserved regions and structural E1, E2, E3, 6K regions and a 3' non-transcriptional region (3'-NTR). The Sindbis-IMB was isolated from the blood of a patient suspected to have encephalitis, and was followed by identification and passage. The virus RNA was extracted from virus supernatant in infected cells and the whole genome was divided into 12 fragments; RT-PCR was then performed to amplify the 12 fragments for complete sequencing. The results showed that the whole genomic sequence of Sindbis-IMB consists of 11 717 bp encoding 3 773 amino acids. Homology comparison with other Sindbis-like isolates demonstrated the highest similarity was the YN87448 with a variation of 1% strain isolated in Yunnan Province and the second highest to the SAAR86 strain with a variation of～1.2%.The nucleotide sequence variations were present in non-structural regions, resulting in amino acids K, E, N, R, H, and L in protein sequences in positions 230, 231, 443,781, 1 582, and 1746 in the new isolation respectively. Furthermore, three additional amino acids--glutamic acid, serine and alanine--were noted in nsp4 terminus as compared to the YN87448 isolate.

Objective To study different test positions on vestibular evoked myogenic potentials (VEMPs) in youth ,and to find a suitable position and provide a guidance for clinical practice .Methods Thirty normal young vol-unteers were tested by vestibular evoked myogenic potentials ,using three different positions :supine with the head held straight up(SHU),supine with the head held up and turned away from the test ear(SHT),sitting with the head turned away from the test(SIT) ,the derivation rate ,latency and amplitude were analyzed .Results The deri-vation rate of SHU ,SHT and SIT were 100% ,100% and 63 .3% ,respectively .The derivation rate ,p13 ,n23 la-tency and p13 n23 inter-latency between SHU and SIT ,and between SHT and SIT had statistical differences (P<0 .05) .No statistical significant differences were found in derivation rate ,p13 ,n23 latency and p13n23 inter-latency between SHU and SHT (P>0 .05) .The amplitude was significantly different among the three positions (P<0 .05) . No statistical significant difference were found in derivation rate ,p13 ,n23 latency ,p13 n23 inter -latency and am-plitude between men and women of the three positions (P> 0 .05) .Conclusion The derivation rate of SHT was 100% with maximum amplitude .SHT is the most recommended position for clinical test in youth .The derivation rate of SHU was 100% ,and no statistical significant difference were found in p13 ,n23 latency and p13n23 inter-la-tency between SHU and SHT (P>0 .05) .SHU can be used in clinical test .SIT is not recommended for using in clinical test .Gender does not affect VEM Ps test .

10.3969/j.issn.1008-9691.2013.03.011