The herpes simplex virus(HSV) Us3 gene encodes a ser/thr protein kinase(PK).As an accessory gene,it plays an important role in the regulation of the apoptosis of infected cells and virus release.Us3-induced alterations in the host cytomorphology are associated with enhanced intercellular virus spread,suggestive of a previously undescribed aspect of alphaherpesvirus spread.

Objective To investigate the function and possible action mechanisms of microRNA hsa-mir-634 in Vero cells.Methods The binding sites for hsa-mir-634 in the 3' UTR of cyclin D1 (CCND1) were predicated by bioinformatics methods.Then,the 3'UTR sequence of CCND1 containing the binding sites for hsamir-634 was amplified by PCR.Site-directed mutagenesis was used to create mutations in the binding sites.The wild and mutant 3' UTR sequences of the CCND1 gene were ligated into the psi-CHECK2 vector separately to construct dual-luciferase reporter vectors,including CHECK2-CCND1 wild,CHECK2-CCND1 mut 1,CHECK2-CCND1 mut 2 and CHECK2-CCND1 mut 3.Then,293T cells were transfected with the four constructed plasmids,and luciferase activity was measured 48 hours after the transfection.Vero cells were transfected with hsa-mir-634 mimics and negative control separately,and harvested after additional culture for different durations; the Vero cells remaining untreated served as the blank control.Subsequently,fluorescence-based quantitative PCR and Western blot were performed to detect the mRNA and protein expressions of CCND1 respectively in,3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt (MTS) assay to evaluate the proliferation of,and flow cytometry to detect the apoptosis in,Vero cells.Results The binding sites for hsa-mir-634 in the 3'UTR of CCND1 were successfully predicated.Sequencing results showed the successful construction of dual-luciferase reporter vectors.As the luciferase assay revealed,the overexpression of hsa-mir-634 could significantly inhibit the CCND1 3'UTR-mediated luciferase activity.Compared with the negative control,the hsamir-634 mimics markedly decreased the protein expression of CCND1,but had no obvious effect on the mRNA expression of CCND1 in Vero cells.The proliferation of Vero cells transfected with hsa-mir-634 mimics was significantly restrained compared with those transfected with the negative control,and the strongest restraining effect was observed on day 4 after the transfection.In addition,the overexpression of hsa-mir-634 also induced the apoptosis of Vero cells,with the apoptosis rate being 8.03％,7.96％ and 17.33％ in the blank control group,negative control group and mimics group respectively.Conclusion Hsa-mir-634 may regulate the proliferation and apoptosis of Vero cells via influencing the expression of CCND1.

Objective To retrospectively analyse the adverse reactions and complications of Graves' disease after thyroid arteries embolization.Methods 41 patients of Graves' disease underwent interventional embolization have been analysed with its adverse reactions and complications. Polyvinyl alcohol or bletilla microspheres and micro coils were used in these patients. Results Laryngopharyngeal and neck pain occurred in all patients. T 3 and T 4 increased in 3 days to one week after the procedure. Thirty of them showed fever. Dystopic embolism happened in two cases with one of transitory hypoparathyroidism. No hypothyroidism or hypoparathyroidism or hoarseness occur during long term follow up.Conclusions The adverse reactions and complications of Graves disease after thyroid arteries embolization may occur. Some of them are preventable and curable.

Objective To study the color doppler image characteristics of thyroid arteries pre and post interventional procedure and to assess the clinical efficacy in Gnaves' disease.Methods 11 from 31 patients diagnosed as Graves' disease undertaken thyroid arteries embolization, were analyzed. Color Doppler sonography was applied to monitor the pre and post procedure thyroid size and diameters of thyroid arteries. Power Doppler was used to detect the Vmax, Vmin, RI and blood flow. Results After thyroid arteries embolization, the size and vascularity of thyroids were reduced. The thyroid arteries showed shrinkage and stoppage blood flow at the embolized site. The changes of RI, blood parameters of Vmax, Vmin and diameters of the thyroid arteries pre and post procedure turned out to be statistically significant for clinical restriction.Conclusion The color Doppler sonography plays an important role for preoperative diagnosis and predicting the prognosis.

BACKGROUND:Currently, inhaled glucocorticoid is still the classic treatment for asthma. Adult stem cell transplantation has made significant progress in a variety of diseases, and it also provides new insights into the treatment of asthma.OBJECTIVE:To summarize the recent advances in the treatment of asthma with adipose-derived stem cells and related adult stem cells, and to discuss the therapeutic safety of adipose-derived stem cells and possible research directions in asthma therapy.METHODS:Relevant articles published from 2001 to 2016 were searched in PubMed, CBM, CNKI, and Wanfang databases. The keywords were (adipose-derived stem cells[All Fields]) OR (adipose stem cells[All Fields]) OR (adipose-derived mesenchymal stem cells[All Fields]) AND (asthma[All Fields]) in English and Chinese, respectively.RESULTS AND CONCLUSION:A total of 125 literatures were initially searched, and finally 54 representative papers were selected. Adipose-derived stem cells may reduce airway inflammation, airway hyperresponsiveness, ease collagen deposition and scar tissue formation, promote neovascularization, and reconstruct damaged airways in the mouse asthma model through immune regulation. It is necessary to understand its treatment mechanism of action deeply and comprehensively and carry out genomic analysis before introduction of adipose-derived stem cells as a conventional clinical treatment. In summary, adipose-derived stem cells may be a therapeutic potential for the treatment of airway allergic inflammatory diseases such as asthma.

Objective To investigate whether or not the pulmonary artery participates in the blood supply of lung cancer and its change of morphology and blood flow in lung cancer. Methods Two different colors of silicone were injected separately into the bronchial and pulmonary arteries of 33 rat models with squamous cell carcinoma of lung. The origin of blood supply of lung cancer and the morphologic change of pulmonary artery were observed under a stereomicroscope. The DSA of bronchial and pulmonary artery were performed simultaneously in 28 patients with lung cancer. Results The pulmonary branch of rat and patients were reduced, thinned and occluded in the affected lung. The pulmonary artery did not form tumor vessel, and pulmonary blood flow and perfusion were reduced or absent in the affected area. Conclusion The pulmonary artery didn′t participate in the blood supply of lung cancer. It is unreasonable to perform transcatheter chemo embolization for lung cancer via pulmonary arteriay.

Objective Up to now, there has been no sure cure for genital herpes (GH), and vaccine seems a most promis-ing approach to the prevention and treatment of herpes simplex virus Ⅱ(HSV-2) infection.In this study, we investigated the feasibili-ty of preparing a dendritic cell ( DC) vaccine modified by the adenovirus-mediated HSV-2 gD gene. Methods We subcloned the HSV-2 gD gene into the vector Shuttle-2 and constructed the recombinant adenovirus pAdeno-HSV-2 gD following identification by en-zyme digestion and DNA sequence analysis .We isolated DCs from the mouse bone marrow , analyzed their phenotypes by flow cytome-try after transfection with the recombinant adenovirus pAdeno-HSV-2 gD, and determined the expression of HSV-2 gD by immunohisto-chemistry, RT-PCR, SDS-PAGE, and Western blot. Results Based on HSV-2 DNA, the corresponding target fragments were am-plified with the gD gene primers.Agarose gel electrophoresis showed the correct size of the PCR product (1182 bp) as predicted.The recombinant adenovirus pAdeno-HSV-2 gD was obtained by transfecting the 293 cells with pAdeno-gD DNA, which had an activity of 4 ×1010 IU/mL.The contents of CD40, CD80, and CD86 were (74.2 ±3.9), (73.9 ±4.1), and (76.1 ±5.5) % in the mature DCs and (81.3 ±3.1), (80.4 ±2.9), and (83.7 ±3.9) % in the pAdeno-HSV-2 gD DCs, significantly increased as compared with those in the immature DCs ([9.7 ±0.5], [7.5 ±1.2], and [5.2 ±1.1] %) (P<0.01).No statistically significant differ-ences were found between the expression of the surface molecules in the pAdeno -HSV-2 gD DCs and that in the cytokine stimulation-in-duced mature DCs (P>0.05).RT-PCR and immunohistochemistry confirmed the expression of HSV-2 gD in DCs.SDS-PAGE and Western blot of the expressed protein showed a new band with an apparent molecular mass corresponding to the predicted size (43000). Conclusion The results of our study have paved the ground for the successful preparation and identification of a dendritic cell vaccine modified by the adenovirus-mediated HSV-2 gD gene.

BACKGROUND: By now, safety of collagenase application is still controvertible and some scholars believed that collagenase might induce the peripheral tissue injury. It attracts much attention in clinic that whether there is nerve injury induced by collagenase chemonucleolysis (CCN) around the injection sites.OBJECTIVE: To observe the effects of collagenase on spinal nerve conduction velocity (NCV) of rats detected with evoked potential method so as to probe into the safetyof collagenase application and further demonstrate the safety of percutaneous intervertebral disc CCN. DESIGN: Randomly grouping design, animal experiment.SETTING: Department of Interventional Radiology, Affiliated First Hospital of Sun Yat-sen University. MATERIALS: The experiment was conducted in the Basic Medical College of Sun Yat-son University from July to September 2002. A total of 57 male SD rats were randomly divided into normal group (n=9), acute sham-operation group (n=10), subacute sham-operation group (n=8), chronic sham-operation group (n=7), acute experimental model group (n=9), subacute experimental model group (n=7) and chronic experimental model group (n=7). METHODS: After being anaesthetized by intraperitoneal injection of continal (45 mg/kg) to separate and identify dorsal root ganglion (DRG),rats in the experimental group were locally dripped with 1 mL of collagenase (300 U/ML) and those in the sham operation group locally dripped with 1 mL of normal saline. Stimulating electrode was placed in the A point of sciatic nerve and recording electrode in the B point of ganglionic central process segment of DRG in L5 nerve root. Evoked potentials A and B were simulated to continuously record latency twice, and the average value was calculated; Distance between A and B were measured and recorded. NCV = distance between A and B / latency. Evoked potential of a segment ofnerve including DRG was measured in the acute group at one hour after administration, in the sub acute group at one week after administration and in the chronic group at one month after administration. MAIN OUTCOME MEASURES: NCV of each group. RESULTS: A total of 57 enrolled animals were involved in the analysis. NCV in normal group, acute experimental group, acute sham-operation group, subacute experimental group, subacute sham-operation group, chronic experimental group, chronic sham-operation group were (45.4±10.7), (43.4±5.9), (46.3±6.5), (52.4±10.4), (49.7±8.1), (46.7±11.0) and (44.6±6.5) m/s respectively. There were no remarkable differences in NCV among all the groups by using one-way analysis of variance (F=1.010,P=0.430); It was showed that there were no marked differences in NCV between each two groups by using multiple comparisons (P=0.336). CONCLUSION: Collagenase at the therapeutic concentration applied in clinical CCN has no remarkable effects on NCV of rat spinal nerve, and to a certain extent, percutaneous intervertebral disc CCN is relatively reliable.

Objective To assess the efficacy and safety of 308 nm excimer laser plus topical pimeevaluated after 15 and 30 times of laser therapy respectively.ResultsExcept for one patient,all patients were able to be evaluated for effiicacy.After 30 times of laser therapy,the response and excellent response rates were 89.6%and 77.1%respectively,in group A，5.0%and 52.1%respectively in group B;both rates were significantly higher in group A than in group B(both P＜0.05).Also,a highler repigmentation rate was obtained in facial lesions in group A compared with group B.Conclusions The 308 nm excimer laser is safe,erective and well-tolerated for vitiligo in childhood,and the combination with topical imecrolimus cream may improve its efficacy in facial vitiligo.

Objective To study the dynamic imaging changes of the uterine leiomyomas before and after uterine arterial embolization (UAE) treatment and to discuss its therapeutic mechanism. Methods Color Doppler senography and both plain and enhanced MR[scanning were performed in 45 patients with uterine leiomyomas before and after UAE. Plain CT scan was performed in all patients after UAE. All the patients were followed up for 3-16 months (average 10±3.5 months). Results In 41 of the total 45 cases, the color Doppler senography showed rich blood flow signals in leiomyomas and myometrium before UAE and no or less blood flow signals in both leiomyomas and myometrium on the first day after UAE. On the seventh day, the blood flow signal was still absent in leiomyomas while it was restored in myometrium, and the same phenomena remained in the first, the third and the twelfth month after UAE. In the other four eases, color Doppler sonography demonstrated blood flow signals inside leiomyomas on the seventh day after UAE and it remained till twelve months after embolization. The embolic agent (Lipiodol) was found in both leiomyomas and myometrium on CT scan for 45 cases on the first day of UAE. CT scan also showed the deposit of the Lipiodol in myometrium, but Lipiodol gradually vanished in leiomyomas at one, three and the twelve months after UAE. The enhancement was apparent in leiomyomas and myometrium on MRI scan in all 45 cases before UAE. The enhancement was found in the myometrium, but not in leiomyomas, on MRI scan in 39 cases 3 months after UAE. The other six cases demonstrated different degrees of enhancement in leinmyomas after embolization. In two cases the detachment of the leiomyomas were observed after embolization and the desquamating materials were pathologicallyproved to be necrotic tissue. The difference in the measuring data about leiomyoma volume between MPI and color Doppler sonography was of no statistical significance (P > 0.05). Conclusion The therapeutic mechanism of UAE for uterine leiomyomas is selectively embolizing the vascular bed of uterus, leading to subsequent necrosis of leiomyomas. The color Doppler sonography should be the fast choice for the dynamic imaging follow-up after UAE.