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.

Objective: To investigate the changing trends in the burden of cardiovascular diseases (CVD) in Zhejiang Province from 1990 to 2019, so as to provide insights into CVD control in the province. Methods: Based on the data of the Global Burden of Disease Study (GBD) 2019 (GBD 2019), the years lived with disability (YLD), years of life lost (YLL), disability-adjusted life years (DALY), YLD rate, YLL rate and DALY rate due to CVD were estimated to assess the burden of CVD in Zhejiang Province from 1990 to 2019. The changing trends in the burden of CVD were using annual percent change (APC) in Zhejiang Province from 1990 to 2019, and compared to China and global regions with different socio-demographic indexes (SDI). Results: The YLL rate (APC=-0.50%, F=22.366, P<0.001) and DALY rate due to CVD (APC=-0.20%, F=5.212, P=0.030) appeared a tendency towards a decline in Zhejiang Province from 1990 to 2019, while the YLD rate showed a tendency towards a rise (APC=1.92%, F=1 117.891, P<0.001). The YLL rate (APC=-1.09%, F=56.871, P<0.001), YLD rate (APC=1.92%, F=922.541, P<0.001) and DALY rate (APC=-0.60%, F=28.974, P<0.001) appeared the same tendency among women relative to among the whole populations, while only the YLD rate showed a clear-cut change among men, appearing a tendency towards a rise (APC=1.82%, F=1 238.423, P<0.001). Both the YLL rate and DALY rate appeared a tendency towards a decline among subjects at all age groups, with the most significant reduction among subjects at ages of 50 to 69 years (YLL rate: APC=-4.40%, F=1 443.608, P<0.001; DALY rate: APC=-3.82%, F=1 519.452, P<0.001). Among all types of CVD, the highest burden was measured in cerebrovascular diseases, with a standardized DALY rate of 1 359.7/105, and the most remarkable reduction in the standardized DALY rate was seen in rheumatic heart diseases. The burden of CVD in Zhejiang Province was lower than that in China and the majority of the world, with consistent changing trends with that in China and global regions with different SDI. Conclusions The overall burden of CVD appeared a tendency towards a decline in Zhejiang Province from 1990 to 2019, which was in agreement with that in China and global regions with different SDI. A remarkable reduction in the burden of CVD was seen among women, residents at ages of 50 to 69 years and cases with rheumatic heart diseases.

Objective To study the effects of semaphorin 5A (SEMA5A) gene silencing by lentivirus?mediated short hairpin RNA(shRNA)on biological activity of malignant melanoma cell line A375. Methods Two pairs of interference sequences for SEMA5A gene(shRNA1 and A375?shRNA2)and a pair of control interference sequences were designed to build lentiviral vectors, which were then transfected into HEK293T cells to gain lentivirus. A375 cells were divided into three groups:experimental group(A375?shRNA1 and A375?shRNA2 cells)transfected with the lentivirus containing shRNA1 or shRNA2, negative control group (A375?con cells) transfected with that containing the control shRNA, and blank control group(A375 cells)receiving no transfection. The A375 cells with stable knockdown of SEMA5A gene expression were screened by puromycin. Subsequently, reverse transcription?PCR and Western?blot analysis were performed to detect mRNA and protein expressions of Semaphorin 5A in these cells, and methyl thiazolyl tetrazolium(MTT)assay was applied to evaluate the growth of cells. The scratch assay and invasion assay were conducted to estimate migration and invasion ability of cells. Results The lentivirus containing the SEMA5A?targeting shRNAs or control shRNA was successfully transfected into A375 cells, and stably transfected cells were gained after puromycin selection. The expressions of semaphorin 5A mRNA and protein in the A375?shRNA2 cells were significantly reduced compared with those in the A375?con and A375 cells(all P < 0.05). MTT assay showed that the growth of A375?shRNA2 cells was significantly slower than that of A375?con and A375 cells(both P<0.05), while there was no significant difference in the growth rate between A375?con and A375 cells(P>0.05). The scratch assay showed that there was no obvious cell migration into the scratch in the experiment group, whereas the scratch was almost covered by cells in the negative control group and blank control group. The invasion assay showed that the number of A375?shRNA2 cells passing through the Transwell chamber was significantly smaller than that of A375 and A375?con cells(both P<0.05), while there was no significant difference between that of A375 and A375?con cells(P > 0.05). Conclusion The silencing of SEMA5A gene by lentivirus?mediated shRNA could effectively down?regulate the expression of semaphorin 5A, and inhibit the growth, invasion and migration of A375 cells.

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.

OBJECTIVETo observe the clinical therapeutic effect of acupuncture massage for primary dysmenorrhea and its influence on prostaglandin F2alpha level in menstrual fluid.

METHODSNinety patients with primary dysmenorrhea were assigned equally to the acupuncture group, the massage group and the control group (oral administrated with Somiton tablet). The treatment course was 3 menstrual cycles. The effect in relieving menalgia was evaluated using numerical rating scale (NRS) before and after treatment and change of PGF2alpha content in menstrual fluid was tested as well.

RESULTSThe NRS score was 0.60 +/- 0.67 in the acupuncture group and 0.53 +/- 0.63 in the massage group respectively, significantly lower than that in the control group, which was 2.30 +/- 1.73 (P < 0.01). The PGF2alpha level after treatment was significantly lower in the acupuncture group (176.58 +/- 17.85 ng/L) and the massage group (171.32 +/- 17.15 ng/L) than that in the control group (220.56 +/- 54.47 ng/L), respectively (P <0.01). There showed no difference in the two aspects between the acupuncture group and the massage group (P >0.05).

CONCLUSIONAcupuncture and massage has a reliable therapeutic effect on primary dysmenorrhea, which is superior to that of Somiton tablet, and its mechanism might be exerted by regulating PGF2alpha level in menstrual fluid.

Acupuncture Therapy ; Decidua ; metabolism ; Dinoprost ; metabolism ; Dysmenorrhea ; therapy ; Female ; Humans ; Massage

Objective To detect and analyze thyroid tumor by two-dimensional sonogram and Doppler parameter, and evaluate the value of high-frequency ultrasound in diagnosing thyroid benign and malignant tumors. Methods The ultrasonic images of 104 thyroid tumor from 80 patients with typical features were collected. Thyroid tumor was classified into benign and malignant nodules, based on the shape, border, or the rear wall echo, echo attenuation loss, internal echo, and microcalcifications in two-dimensional sonogram and systolic blood peak velocity (Vmax) and resistant index (RI) in Doppler examination. The expected results of high frequency ultrasound were compared with pathological results on consistency and error rate. Results Prediction of benign tumor by high frequency ultrasound was 66, and pathology 61, consistency rate of the two was 92.4%. Prediction of malignant lesions was 38, and pathological examination 32, consistency rate of the two was 84.2%. The total coincidence rate was 89.4%(93/104) and the error rate was 11.6%( 11/104). Conclusions The typical features of thyroid tumor on high-frequency ultrasound are helpful in diagnosis of benign or malignant nodules, which is valuable in guiding clinical treatment.

In this study, 31 Notopterygium incisum populations were analyzed using ITS sequences to investigate the genetic structure. The results showed that: the ITS region ranged in size from 634 to 635 bp and base composition was with high G + C content of 57.8%. Thirty-one polymorphic sites were detected from 402 sequences of 31 populations of N. incisum, and the proportion of polymorphic sites was 4.88%, in which parsimony informative sites were up to 12. And 31 haplotypes were identified based on these polymorphic sites. Molecular variance analysis (AMOVA) indicated that high genetic differentiation (57%) existed among population, and gene flow was low (N(m) = 0.38) among populations. Phylogenetic relationships of 31 haplotypes were analyzed using NJ method with N. forbesiias an out-group. Phylogenetic analysis showed that 31 haplotypes from different populations mixed together and did not form distinct geographically separated clades.
Apiaceae ; classification ; genetics ; Base Sequence ; China ; DNA, Intergenic ; genetics ; Gene Flow ; Genetic Variation ; Molecular Sequence Data ; Phylogeny

Arrival of qi, an important component of traditional theory of acupuncture and moxibustion, is the key factor in determining clinical therapeutic effect of acupuncture. The authors summarize the indication of qi-arrival, correlativity of qi-arrival with clinical therapeutic effect and its action mechanism through reviewing modern clinical research. At present, there is more subjective in the study on qi-arrival, so the key problem in research on qi-arrival is how the qi-arrival state is measured by an objective quantization standard. The cerebral function imaging, a new technique, may be used in objectification research on qi-arrival.
Acupuncture Points ; Acupuncture Therapy ; Humans ; Qi ; Research

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.

Herpes simplex virusⅠ(HSVⅠ) regulating the pathway of transcription and translation modify in host cell is a very systematic and complicate system. A clear understanding of the concrete mechanisms of infection will greatly help to comprehend the virus replication and the interaction with the host cell. By the analysis of 2-DE, the heterogeneous nuclear ribonucleoprotein H2 in human fetal liver cell represent distinction after the HSVⅠinfection.Utilization of Northern blot and Western blot technologies verified the expression of hnRNP H2 in different stage of virus infection is varied.